Boron-containing small molecules

ABSTRACT

This invention relates to compounds useful for treating fungal infections, more specifically topical treatment of onychomycosis and/or cutaneous fungal infections. This invention is directed to compounds that are active against fungi and have properties that allow the compound, when placed in contact with a patient, to reach the particular part of the skin, nail, hair, claw or hoof infected by the fungus. In particular the present compounds have physiochemical properties that facilitate penetration of the nail plate.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a continuation of U.S. patent application Ser. No.13/224,252 filed Sep. 1, 2011, now U.S. Pat. No. 8,440,642, which is acontinuation of Ser. No. 12/507,010 filed Jul. 21, 2009, now U.S. Pat.No. 8,039,451, which is a continuation of Ser. No. 11/357,687 filed Feb.16, 2006, now U.S. Pat. No. 7,582,621, which claims priority to60/654,060 filed Feb. 16, 2005, the full disclosure of which isincorporated herein in its entirety for all purposes.

BACKGROUND FOR THE INVENTION

Infections of the nail and hoof, known as ungual and/or periungualinfections, pose serious problems in dermatology. These ungual and/orperiungual can be caused by sources such as fungi, viruses, yeast,bacteria and parasites. Onychomycosis is an example of these seriousungual and/or periungual infections and is caused by at least onefungus. Current treatment for ungual and/or periungual infectionsgenerally falls into three categories: systemic administration ofmedicine; surgical removal of all or part of the nail or hoof followedby topical treatment of the exposed tissue; or topical application ofconventional creams, lotions, gels or solutions, frequently includingthe use of bandages to keep these dosage forms in place on the nail orhoof. All of these approaches have major drawbacks. The followingdiscussion is particularly directed to drawbacks associated with currenttreatment of ungual and/or periungual antifungal infections.

Long term systemic (oral) administration of an antifungal agent for thetreatment of onychomycosis is often required to produce a therapeuticeffect in the nail bed. For example, oral treatment with the antifungalcompound ketoconozole typically requires administration of 200 to 400mg/day for 6 months before any significant therapeutic benefit isrealized. Such long term, high dose systemic therapy can havesignificant adverse effects. For example, ketoconozole has been reportedto have liver toxicity effects and reduces testosterone levels in blooddue to adverse effects on the testes. Patient compliance is a problemwith such long term therapies especially those which involve seriousadverse effects. Moreover, this type of long term oral therapy isinconvenient in the treatment of a horse or other ruminants afflictedwith fungal infections of the hoof. Accordingly, the risks associatedwith parenteral treatments generate significant disincentive againsttheir use and considerable patient non-compliance.

Surgical removal of all or part of the nail followed by topicaltreatment also has severe drawbacks. The pain and discomfort associatedwith the surgery and the undesirable cosmetic appearance of the nail ornail bed represent significant problems, particularly for femalepatients or those more sensitive to physical appearance. Generally, thistype of treatment is not realistic for ruminants such as horses.

Topical therapy has significant problems too. Topical dosage forms suchas creams, lotions, gels etc., can not keep the drug in intimate contactwith the infected area for therapeutically effective periods of time.Bandages have been used to hold drug reservoirs in place in an attemptto enhance absorption of the pharmaceutical agent. However the bandagesare thick, awkward, troublesome and generally lead to poor patientcompliance.

Hydrophilic and hydrophobic film forming topical antifungal solutionshave also been developed. These dosage forms provide improved contactbetween the drug and the nail, but the films are not occlusive. Topicalformulations for fungal infection treatment have largely tried todeliver the drug to the target site (an infected nail bed) by diffusionacross or through the nail.

Nail is more like hair than stratum corneum with respect to chemicalcomposition and permeability. Nitrogen is the major component of thenail attesting to the nail's proteinaceous nature. The total lipidcontent of mature nail is 0.1-1.0%, while the stratum corneum lipid isabout 10% w/w. The nail is 100-200 times thicker than the stratumcorneum and has a very high affinity and capacity for binding andretaining antifungal drugs. Consequently little if any drug penetratesthrough the nail to reach the target site. Because of these reasonstopical therapy for fungal infections have generally been ineffective.

Compounds known as penetration or permeation enhancers are well known inthe art to produce an increase in the permeability of skin or other bodymembranes to a pharmacologically active agent. The increasedpermeability allows an increase in the rate at which the drug permeatesthrough the skin and enters the blood stream. Penetration enhancers havebeen successful in overcoming the impermeability of pharmaceuticalagents through the skin. However, the thin stratum corneum layer of theskin, which is about 10 to 15 cells thick and is formed naturally bycells migrating toward the skin surface from the basal layer, has beeneasier to penetrate than nails. Moreover, known penetration enhancershave not proven to be useful in facilitating drug migration through thenail tissue.

Antimicrobial compositions for controlling bacterial and fungalinfections comprising a metal chelate of 8-hydroxyquinoline and an alkylbenzene sulfonic acid have been shown to be efficacious due to theincreased ability of the oleophilic group to penetrate the lipoid layersof micro-cells. The compounds however, do not effectively increase theability to carry the pharmaceutically active antifungal through thecornified layer or stratum corneum of the skin. U.S. Pat. No. 4,602,011,West et al., Jul. 22, 1986; U.S. Pat. No. 4,766,113, West et al., Aug.23, 1988.

Therefore, there is a need in the art for compounds which caneffectively penetrate the nail. There is also need in the art forcompounds which can effectively treat ungual and/or periungualinfections. These and other needs are addressed by the currentinvention.

SUMMARY OF THE INVENTION

In a first aspect, the invention provides a compound having a structureaccording to Formula I:

wherein B is boron. R^(1a) is a member selected from a negative charge,a salt counterion, H, substituted or unsubstituted alkyl, substituted orunsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl,substituted or unsubstituted heterocycloalkyl, substituted orunsubstituted aryl, and substituted or unsubstituted heteroaryl. M1 is amember selected from oxygen, sulfur and NR^(2a). R^(2a) is a memberselected from H, substituted or unsubstituted alkyl, substituted orunsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl,substituted or unsubstituted heterocycloalkyl, substituted orunsubstituted aryl, and substituted or unsubstituted heteroaryl. J1 is amember selected from (CR^(3a)R^(4a))_(n1) and CR^(5a). R^(3a), R^(4a),and R^(5a) are members independently selected from H, OH, NH₂, SH,substituted or unsubstituted alkyl, substituted or unsubstitutedheteroalkyl, substituted or unsubstituted cycloalkyl, substituted orunsubstituted heterocycloalkyl, substituted or unsubstituted aryl, andsubstituted or unsubstituted heteroaryl. The index n1 is an integerselected from 0 to 2. W1 is a member selected from C═O (carbonyl),(CR^(6a)R^(7a))_(m1) and CR^(8a). R^(6a), R^(7a), and R^(8a) are membersindependently selected from H, OH, NH₂, SH, substituted or unsubstitutedalkyl, substituted or unsubstituted heteroalkyl, substituted orunsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl,substituted or unsubstituted aryl, and substituted or unsubstitutedheteroaryl. The index m1 is an integer selected from 0 and 1. A1 is amember selected from CR^(9a) and N. D1 is a member selected fromCR^(10a) and N. E1 is a member selected from CR^(11a) and N. G1 is amember selected from CR^(12a) and N. R^(9a), R^(10a), R^(11a) andR^(12a) are members independently selected from H, OH, NH₂, SH,substituted or unsubstituted alkyl, substituted or unsubstitutedheteroalkyl, substituted or unsubstituted cycloalkyl, substituted orunsubstituted heterocycloalkyl, substituted or unsubstituted aryl, andsubstituted or unsubstituted heteroaryl. The combination of nitrogens(A1+D1+E1+G1) is an integer selected from 0 to 3. A member selected fromR^(3a), R^(4a) and R^(5a) and a member selected from R^(6a), R^(7a) andR^(8a), together with the atoms to which they are attached, areoptionally joined to form a 4 to 7 membered ring. R^(3a) and R^(4a),together with the atoms to which they are attached, are optionallyjoined to form a 4 to 7 membered ring. R^(6a) and R^(7a), together withthe atoms to which they are attached, are optionally joined to form a 4to 7 membered ring. R^(9a) and R^(10a), together with the atoms to whichthey are attached, are optionally joined to form a 4 to 7 membered ring.R^(10a) and R^(11a), together with the atoms to which they are attached,are optionally joined to form a 4 to 7 membered ring. R^(11a) andR^(12a), together with the atoms to which they are attached, areoptionally joined to form a 4 to 7 membered ring. The aspect has theproviso that when M1 is oxygen, W1 is a member selected from(CR^(3a)R^(4a))_(n1), wherein n1 is 0, J1 is a member selected from(CR^(6a)R^(7a))_(m1), wherein m1 is 1, A1 is CR^(9a), D1 is CR^(10a), E1is CR^(11a), G1 is CR^(12a), then R^(9a) is not halogen, methyl, ethyl,or optionally joined with R^(10a) to a form phenyl ring; R^(10a) is notunsubstituted phenoxy, C(CH₃)₃, halogen, CF₃, methoxy, ethoxy, oroptionally joined with R^(9a) to form a phenyl ring; R^(11a) is nothalogen or optionally joined with R^(10a) to form a phenyl ring; andR^(12a) is not halogen. The aspect has the further proviso that when M1is oxygen, W1 is a member selected from (CR^(3a)R^(4a))_(n1), wherein n1is 0, J1 is a member selected from (CR^(6a)R^(7a))_(m1), wherein m1 is1, A1 is CR^(9a), D1 is CR^(10a), E1 is CR^(11a), G1 is CR^(12a), thenneither R^(6a) nor R^(7a) are halophenyl. The aspect has the furtherproviso that when M1 is oxygen, W1 is a member selected from(CR^(3a)R^(4a))_(n1), wherein n1 is 0, J1 is a member selected from(CR^(6a)R^(7a))_(m1), wherein m1 is 1, A1 is CR^(9a), D1 is CR^(10a), E1is CR^(11a), G1 is CR^(12a), and R^(9a), R^(10a) and R^(11a) are H, thenR^(6a), R^(7a) and R^(12a) are not H. The aspect has the further provisothat when M1 is oxygen wherein n1 is 1, J1 is a member selected from(CR^(6a)R^(7a))_(m1), wherein m1 is 0, A1 is CR^(9a), D1 is CR^(10a), E1is CR^(11a), G1 is CR^(12a), R^(9a) is H, R^(10a) is H, R^(11a) is H,R^(6a) is H, R^(7a) is H, R^(12a) is H, then W1 is not C═O (carbonyl).The aspect has the further proviso that when M1 is oxygen, W1 isCR^(5a), J1 is CR^(8a), A1 is CR^(9a), D1 is CR^(10a), E1 is CR^(11a),G1 is CR^(12a), R^(6a), R^(7a), R^(9a), R^(10a), R^(11a) and R^(12a) areH, then R^(5a) and R^(8a), together with the atoms to which they areattached, do not form a phenyl ring.

In a second aspect, the invention provides a pharmaceutical formulationcomprising (a) a pharmaceutically acceptable excipient; and (b) acompound having a structure according to Formula II:

wherein B is boron. R^(1b) is a member selected from a negative charge,a salt counterion, H, substituted or unsubstituted alkyl, substituted orunsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl,substituted or unsubstituted heterocycloalkyl, substituted orunsubstituted aryl, and substituted or unsubstituted heteroaryl. M2 is amember selected from oxygen, sulfur and NR^(2b). R^(2b) is a memberselected from H, substituted or unsubstituted alkyl, substituted orunsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl,substituted or unsubstituted heterocycloalkyl, substituted orunsubstituted aryl, and substituted or unsubstituted heteroaryl. J2 is amember selected from (CR^(3b)R^(4b))_(n2) and CR^(5b). R^(3b), R^(4b),and R^(5b) are members independently selected from H, OH, NH₂, SH,substituted or unsubstituted alkyl, substituted or unsubstitutedheteroalkyl, substituted or unsubstituted cycloalkyl, substituted orunsubstituted heterocycloalkyl, substituted or unsubstituted aryl, andsubstituted or unsubstituted heteroaryl. The index n2 is an integerselected from 0 to 2. W2 is a member selected from C═O (carbonyl),(CR^(6b)R^(7b))_(m2) and CR^(8b). R^(6b), R^(7b), and R^(8b) are membersindependently selected from H, OH, NH₂, SH, substituted or unsubstitutedalkyl, substituted or unsubstituted heteroalkyl, substituted orunsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl,substituted or unsubstituted aryl, and substituted or unsubstitutedheteroaryl. The index m2 is an integer selected from 0 and 1. A2 is amember selected from CR^(9b) and N. D2 is a member selected fromCR^(10b) and N. E2 is a member selected from CR^(11b) and N. G2 is amember selected from CR^(12b) and N. R^(9b), R^(10b), R^(11b) andR^(12b) are members independently selected from H, OH, NH₂, SH,substituted or unsubstituted alkyl, substituted or unsubstitutedheteroalkyl, substituted or unsubstituted cycloalkyl, substituted orunsubstituted heterocycloalkyl, substituted or unsubstituted aryl, andsubstituted or unsubstituted heteroaryl. The combination of nitrogens(A2+D2+E2+G2) is an integer selected from 0 to 3. A member selected fromR^(3b), R^(4b) and R^(5b) and a member selected from R^(6b), R^(7b) andR^(8b), together with the atoms to which they are attached, areoptionally joined to form a 4 to 7 membered ring. R^(3b) and R^(4b),together with the atoms to which they are attached, are optionallyjoined to form a 4 to 7 membered ring. R^(6b) and R^(7b), together withthe atoms to which they are attached, are optionally joined to form a 4to 7 membered ring. R^(9b) and R^(10b), together with the atoms to whichthey are attached, are optionally joined to form a 4 to 7 membered ring.R^(10b) and R^(11b), together with the atoms to which they are attached,are optionally joined to form a 4 to 7 membered ring. R^(11b) andR^(12b), together with the atoms to which they are attached, areoptionally joined to form a 4 to 7 membered ring.

In another aspect, the invention provides a method of killing amicroorganism, comprising contacting the microorganism with atherapeutically effective amount of a compound of the invention.

In another aspect, the invention provides a method of inhibitingmicroorganism growth, comprising contacting the microorganism with atherapeutically effective amount of a compound of the invention.

In another aspect, the invention provides a method of treating aninfection in an animal, comprising administering to the animal atherapeutically effective amount of a compound of the invention.

In another aspect, the invention provides a method of preventing aninfection in an animal, comprising administering to the animal atherapeutically effective amount of a compound of the invention.

In another aspect, the invention provides a method of treating asystemic infection or an ungual or periungual infection in a human,comprising administering to the animal a therapeutically effectiveamount of a compound of the invention.

In another aspect, the invention provides a method of treatingonychomycosis in a human, comprising administering to the animal atherapeutically effective amount of a compound of the invention.

In another aspect, the invention provides a method of synthesizing acompound of the invention.

In another aspect, the invention provides a method of delivering acompound from the dorsal layer of the nail plate to the nail bed. Themethod comprises contacting said cell with a compound capable ofpenetrating the nail plate, under conditions sufficient to penetratesaid nail plate, and thereby delivering the compound. The compound has amolecular weight of between about 100 and about 200 Da. The compoundalso has a log P value of between about 1.0 and about 2.6. The compoundhas a water solubility between about 0.1 mg/mL and 1.0 g/mLoctanol/saturated water.

BRIEF DESCRIPTION OF THE DRAWINGS

FIGS. 1A-1C is a table of minimum inhibitory concentration (MIC) data ofCBO against various fungi.

FIG. 2A displays minimum inhibitory concentration (MIC) for C10,ciclopirox, terbinafine, fluconazole and itraconazole (comparator drugs)against 19 test strains of fungi.

FIG. 2B displays minimum fungicidal concentration (MFC) for C10,ciclopirox, terbinafine and itraconazole (comparator drugs) against 2test strains of fungi.

FIG. 3 displays a comparison of Normalized C10 and Ciclopirox Equivalentin Each Part of Nail Plate Samples after 14-day Treatment.

FIG. 4 displays a comparison of C10 and Ciclopirox Equivalent in CottonBall Supporting Bed Samples after 14-day Treatment.

FIG. 5 displays the results of a placebo for C10 (50:50 propylene glycoland ethyl acetate) applied per day over five days. Full carpet growth ofthe organism T. rubrum was observed.

FIG. 6 displays the results of a 40 μL/cm² aliquot of C10 10% w/vsolution applied per day over five days. Zones of inhibition (in theorder of the cells shown in the figure) of 100%, 67%, 46%, 57%, 38% and71% were observed for the growth of T. rubrum. Green arrow indicates themeasurement of zone of inhibition.

FIG. 7 displays the results of a 40 μL/cm² aliquot of C10 10% w/vsolution applied per day over five days. Zones of inhibition (in theorder of the cells shown in the figure) of 74%, 86%, 100%, 82%, 100% and84% were observed for the growth of T. rubrum.

FIG. 8 displays the results of a 40 μL/cm² aliquot of 8% ciclopirox inw/w commercial lacquer applied per day over five days. No zone ofinhibition observed; full carpet growth of T. rubrum.

FIG. 9 displays the results of a 40 μL/cm² aliquot of 5% amorolfine w/vin commercial lacquer applied per day over five days. No zone ofinhibition observed; full carpet growth of T. rubrum.

DETAILED DESCRIPTION OF THE INVENTION

I. Definitions and Abbreviations

The abbreviations used herein generally have their conventional meaningwithin the chemical and biological arts.

“Compound of the invention,” as used herein refers to the compoundsdiscussed herein, pharmaceutically acceptable salts and prodrugs ofthese compounds.

MIC, or minimum inhibitory concentration, is the point where compoundstops more than 90% of cell growth relative to an untreated control.

Where substituent groups are specified by their conventional chemicalformulae, written from left to right, they equally encompass thechemically identical substituents, which would result from writing thestructure from right to left, e.g., —CH₂O— is intended to also recite—OCH₂—.

The term “poly” as used herein means at least 2. For example, apolyvalent metal ion is a metal ion having a valency of at least 2.

“Moiety” refers to the radical of a molecule that is attached to anothermoiety.

The symbol

, whether utilized as a bond or displayed perpendicular to a bond,indicates the point at which the displayed moiety is attached to theremainder of the molecule.

The term “alkyl,” by itself or as part of another substituent, means,unless otherwise stated, a straight or branched chain, or cyclichydrocarbon radical, or combination thereof, which may be fullysaturated, mono- or polyunsaturated and can include di- and multivalentradicals, having the number of carbon atoms designated (i.e. C₁-C₁₀means one to ten carbons). Examples of saturated hydrocarbon radicalsinclude, but are not limited to, groups such as methyl, ethyl, n-propyl,isopropyl, n-butyl, t-butyl, isobutyl, sec-butyl, cyclohexyl,(cyclohexyl)methyl, cyclopropylmethyl, homologs and isomers of, forexample, n-pentyl, n-hexyl, n-heptyl, n-octyl, and the like. Anunsaturated alkyl group is one having one or more double bonds or triplebonds. Examples of unsaturated alkyl groups include, but are not limitedto, vinyl, 2-propenyl, crotyl, 2-isopentenyl, 2-(butadienyl),2,4-pentadienyl, 3-(1,4-pentadienyl), ethynyl, 1- and 3-propynyl,3-butynyl, and the higher homologs and isomers. The term “alkyl,” unlessotherwise noted, is also meant to include those derivatives of alkyldefined in more detail below, such as “heteroalkyl.” Alkyl groups thatare limited to hydrocarbon groups are termed “homoalkyl”.

The term “alkylene” by itself or as part of another substituent means adivalent radical derived from an alkane, as exemplified, but notlimited, by —CH₂CH₂CH₂CH₂—, and further includes those groups describedbelow as “heteroalkylene.” Typically, an alkyl (or alkylene) group willhave from 1 to 24 carbon atoms, with those groups having 10 or fewercarbon atoms being preferred in the present invention. A “lower alkyl”or “lower alkylene” is a shorter chain alkyl or alkylene group,generally having eight or fewer carbon atoms.

The terms “alkoxy,” “alkylamino” and “alkylthio” (or thioalkoxy) areused in their conventional sense, and refer to those alkyl groupsattached to the remainder of the molecule via an oxygen atom, an aminogroup, or a sulfur atom, respectively.

The term “heteroalkyl,” by itself or in combination with another term,means, unless otherwise stated, a stable straight or branched chain, orcyclic hydrocarbon radical, or combinations thereof, consisting of thestated number of carbon atoms and at least one heteroatom. In anexemplary embodiment, the heteroatoms can be selected from the groupconsisting of B, O, N and S, and wherein the nitrogen and sulfur atomsmay optionally be oxidized and the nitrogen heteroatom may optionally bequaternized. The heteroatom(s) B, O, N and S may be placed at anyinterior position of the heteroalkyl group or at the position at whichthe alkyl group is attached to the remainder of the molecule. Examplesinclude, but are not limited to, —CH₂—CH₂—O—CH₃, —CH₂—CH₂—NH—CH₃,—CH₂—CH₂—N(CH₃)—CH₃, —CH₂—S—CH₂—CH₃, —CH₂—CH₂, —S(O)—CH₃,—CH₂—CH₂—S(O)₂—CH₃, —CH═CH—O—CH₃, —CH₂—CH═N—OCH₃, and —CH═CH—N(CH₃)—CH₃.Up to two heteroatoms may be consecutive, such as, for example,—CH₂—NH—OCH₃. Similarly, the term “heteroalkylene” by itself or as partof another substituent means a divalent radical derived fromheteroalkyl, as exemplified, but not limited by, —CH₂—CH₂—S—CH₂—CH₂— and—CH₂—S—CH₂—CH₂—NH—CH₂—. For heteroalkylene groups, heteroatoms can alsooccupy either or both of the chain termini (e.g., alkyleneoxy,alkylenedioxy, alkyleneamino, alkylenediamino, and the like). Stillfurther, for alkylene and heteroalkylene linking groups, no orientationof the linking group is implied by the direction in which the formula ofthe linking group is written. For example, the formula —C(O)₂R′—represents both —C(O)₂R′— and —R′C(O)₂—.

The terms “cycloalkyl” and “heterocycloalkyl”, by themselves or incombination with other terms, represent, unless otherwise stated, cyclicversions of “alkyl” and “heteroalkyl”, respectively. Additionally, forheterocycloalkyl, a heteroatom can occupy the position at which theheterocycle is attached to the remainder of the molecule. Examples ofcycloalkyl include, but are not limited to, cyclopentyl, cyclohexyl,1-cyclohexenyl, 3-cyclohexenyl, cycloheptyl, and the like. Examples ofheterocycloalkyl include, but are not limited to,1-(1,2,5,6-tetrahydropyridyl), 1-piperidinyl, 2-piperidinyl,3-piperidinyl, 4-morpholinyl, 3-morpholinyl, tetrahydrofuran-2-yl,tetrahydrofuran-3-yl, tetrahydrothien-2-yl, tetrahydrothien-3-yl,1-piperazinyl, 2-piperazinyl, and the like.

The terms “halo” or “halogen,” by themselves or as part of anothersubstituent, mean, unless otherwise stated, a fluorine, chlorine,bromine, or iodine atom. Additionally, terms such as “haloalkyl,” aremeant to include monohaloalkyl and polyhaloalkyl. For example, the term“halo(C₁-C₄)alkyl” is mean to include, but not be limited to,trifluoromethyl, 2,2,2-trifluoroethyl, 4-chlorobutyl, 3-bromopropyl, andthe like.

The term “aryl” means, unless otherwise stated, a polyunsaturated,aromatic, substituent that can be a single ring or multiple rings(preferably from 1 to 3 rings), which are fused together or linkedcovalently. The term “heteroaryl” refers to aryl groups (or rings) thatcontain from one to four heteroatoms. In an exemplary embodiment, theheteroatom is selected from B, N, O, and S, wherein the nitrogen andsulfur atoms are optionally oxidized, and the nitrogen atom(s) areoptionally quaternized. A heteroaryl group can be attached to theremainder of the molecule through a heteroatom. Non-limiting examples ofaryl and heteroaryl groups include phenyl, 1-naphthyl, 2-naphthyl,4-biphenyl, 1-pyrrolyl, 2-pyrrolyl, 3-pyrrolyl, 3-pyrazolyl,2-imidazolyl, 4-imidazolyl, pyrazinyl, 2-oxazolyl, 4-oxazolyl,2-phenyl-4-oxazolyl, 5-oxazolyl, 3-isoxazolyl, 4-isoxazolyl,5-isoxazolyl, 2-thiazolyl, 4-thiazolyl, 5-thiazolyl, 2-furyl, 3-furyl,2-thienyl, 3-thienyl, 2-pyridyl, 3-pyridyl, 4-pyridyl, 2-pyrimidyl,4-pyrimidyl, 5-benzothiazolyl, purinyl, 2-benzimidazolyl, 5-indolyl,1-isoquinolyl, 5-isoquinolyl, 2-quinoxalinyl, 5-quinoxalinyl,3-quinolyl, and 6-quinolyl. Substituents for each of the above notedaryl and heteroaryl ring systems are selected from the group ofacceptable substituents described below.

For brevity, the term “aryl” when used in combination with other terms(e.g., aryloxy, arylthioxy, arylalkyl) includes both aryl and heteroarylrings as defined above. Thus, the term “arylalkyl” is meant to includethose radicals in which an aryl group is attached to an alkyl group(e.g., benzyl, phenethyl, pyridylmethyl and the like) including thosealkyl groups in which a carbon atom (e.g., a methylene group) has beenreplaced by, for example, an oxygen atom (e.g., phenoxymethyl,2-pyridyloxymethyl, 3-(1-naphthyloxy)propyl, and the like).

Each of the above terms (e.g., “alkyl,” “heteroalkyl,” “aryl” and“heteroaryl”) are meant to include both substituted and unsubstitutedforms of the indicated radical. Preferred substituents for each type ofradical are provided below.

Substituents for the alkyl and heteroalkyl radicals (including thosegroups often referred to as alkylene, alkenyl, heteroalkylene,heteroalkenyl, alkynyl, cycloalkyl, heterocycloalkyl, cycloalkenyl, andheterocycloalkenyl) are generically referred to as “alkyl groupsubstituents,” and they can be one or more of a variety of groupsselected from, but not limited to: —OR′, ═O, ═NR′, ═N—OR′, —NR′R″, —SR′,-halogen, —OC(O)R′, —C(O)R′, —CO₂R′, —CONR′R″, —OC(O)NR′R″, —NR″C(O)R′,—NR′—C(O)NR″R′″, —NR″C(O)₂R′, —NR—C(NR′R″R′″)═NR″″, —NR—C(NR′R″)═NR′″,—S(O)R′, —S(O)₂R′, —S(O)₂NR′R″, —NRSO₂R′, —CN and —NO₂ in a numberranging from zero to (2 m′+1), where m′ is the total number of carbonatoms in such radical. R′, R″, R′″ and R′ each preferably independentlyrefer to hydrogen, substituted or unsubstituted heteroalkyl, substitutedor unsubstituted aryl, e.g., aryl substituted with 1-3 halogens,substituted or unsubstituted alkyl, alkoxy or thioalkoxy groups, orarylalkyl groups. When a compound of the invention includes more thanone R group, for example, each of the R groups is independently selectedas are each R′, R″, R′″ and R″″ groups when more than one of thesegroups is present. When R′ and R″ are attached to the same nitrogenatom, they can be combined with the nitrogen atom to form a 5-, 6-, or7-membered ring. For example, —NR′R″ is meant to include, but not belimited to, 1-pyrrolidinyl and 4-morpholinyl. From the above discussionof substituents, one of skill in the art will understand that the term“alkyl” is meant to include groups including carbon atoms bound togroups other than hydrogen groups, such as haloalkyl (e.g., —CF₃ and—CH₂CF₃) and acyl (e.g., —C(O)CH₃, —C(O)CF₃, —C(O)CH₂OCH₃, and thelike).

Similar to the substituents described for the alkyl radical,substituents for the aryl and heteroaryl groups are generically referredto as “aryl group substituents.” The substituents are selected from, forexample: halogen, —OR′, ═O, ═NR′, ═N—OR′, —NR′R″, —SR′, -halogen,—OC(O)R′, —C(O)R′, —CO₂R′, —CONR′R″, —OC(O)NR′R″, —NR″C(O)R′,—NR′—C(O)NR″R′″, —NR″C(O)₂R′, —NR—C(NR′R″R′″)═NR″″, —NR—C(NR′R″)═NR′″,—S(O)R′, —S(O)₂R′, —S(O)₂NR′R″, —NRSO₂R′, —CN and —NO₂, —R′, —N₃,—CH(Ph)₂, fluoro(C₁-C₄)alkoxy, and fluoro(C₁-C₄)alkyl, in a numberranging from zero to the total number of open valences on the aromaticring system; and where R′, R″, R′″ and R″″ are preferably independentlyselected from hydrogen, substituted or unsubstituted alkyl, substitutedor unsubstituted heteroalkyl, substituted or unsubstituted aryl andsubstituted or unsubstituted heteroaryl. When a compound of theinvention includes more than one R group, for example, each of the Rgroups is independently selected as are each R′, R″, R′″ and R″″ groupswhen more than one of these groups is present.

Two of the substituents on adjacent atoms of the aryl or heteroaryl ringmay optionally be replaced with a substituent of the formula-T-C(O)—(CRR′)_(q)-U-, wherein T and U are independently —NR—, —O—,—CRR′— or a single bond, and q is an integer of from 0 to 3.Alternatively, two of the substituents on adjacent atoms of the aryl orheteroaryl ring may optionally be replaced with a substituent of theformula -A-(CH₂)_(r)-B-, wherein A and B are independently —CRR′—, —O—,—NR—, —S—, —S(O)—, —S(O)₂—, —S(O)₂NR′— or a single bond, and r is aninteger of from 1 to 4. One of the single bonds of the new ring soformed may optionally be replaced with a double bond. Alternatively, twoof the substituents on adjacent atoms of the aryl or heteroaryl ring mayoptionally be replaced with a substituent of the formula—(CRR′)_(s)—X—(CR″R′″)_(d)—, where s and d are independently integers offrom 0 to 3, and X is —O—, —NR′—, —S—, —S(O)—, —S(O)₂—, or —S(O)₂NR′—.The substituents R, R′, R″ and R′″ are preferably independently selectedfrom hydrogen or substituted or unsubstituted (C₁-C₆)alkyl.

“Ring” as used herein means a substituted or unsubstituted cycloalkyl,substituted or unsubstituted heterocycloalkyl, substituted orunsubstituted aryl, or substituted or unsubstituted heteroaryl. A ringincludes fused ring moieties. The number of atoms in a ring is typicallydefined by the number of members in the ring. For example, a “5- to7-membered ring” means there are 5 to 7 atoms in the encirclingarrangement. The ring optionally included a heteroatom. Thus, the term“5- to 7-membered ring” includes, for example pyridinyl and piperidinyl.The term “ring” further includes a ring system comprising more than one“ring”, wherein each “ring” is independently defined as above.

As used herein, the term “heteroatom” includes atoms other than carbon(C) and hydrogen (H). Examples include oxygen (O), nitrogen (N) sulfur(S), silicon (Si), germanium (Ge), aluminum (Al) and boron (B).

The symbol “R” is a general abbreviation that represents a substituentgroup that is selected from substituted or unsubstituted alkyl,substituted or unsubstituted heteroalkyl, substituted or unsubstitutedaryl, substituted or unsubstituted heteroaryl, substituted orunsubstituted cycloalkyl and substituted or unsubstitutedheterocycloalkyl groups.

By “effective” amount of a drug, formulation, or permeant is meant asufficient amount of a active agent to provide the desired local orsystemic effect. A “Topically effective,” “Cosmetically effective,”“pharmaceutically effective,” or “therapeutically effective” amountrefers to the amount of drug needed to effect the desired therapeuticresult.

“Topically effective” refers to a material that, when applied to theskin, nail, hair, claw or hoof produces a desired pharmacological resulteither locally at the place of application or systemically as a resultof transdermal passage of an active ingredient in the material.

“Cosmetically effective” refers to a material that, when applied to theskin, nail, hair, claw or hoof, produces a desired cosmetic resultlocally at the place of application of an active ingredient in thematerial.

The term “pharmaceutically acceptable salts” is meant to include saltsof the compounds of the invention which are prepared with relativelynontoxic acids or bases, depending on the particular substituents foundon the compounds described herein. When compounds of the presentinvention contain relatively acidic functionalities, base addition saltscan be obtained by contacting the neutral form of such compounds with asufficient amount of the desired base, either neat or in a suitableinert solvent. Examples of pharmaceutically acceptable base additionsalts include sodium, potassium, calcium, ammonium, organic amino, ormagnesium salt, or a similar salt. When compounds of the presentinvention contain relatively basic functionalities, acid addition saltscan be obtained by contacting the neutral form of such compounds with asufficient amount of the desired acid, either neat or in a suitableinert solvent. Examples of pharmaceutically acceptable acid additionsalts include those derived from inorganic acids like hydrochloric,hydrobromic, nitric, carbonic, monohydrogencarbonic, phosphoric,monohydrogenphosphoric, dihydrogenphosphoric, sulfuric,monohydrogensulfuric, hydriodic, or phosphorous acids and the like, aswell as the salts derived from relatively nontoxic organic acids likeacetic, propionic, isobutyric, maleic, malonic, benzoic, succinic,suberic, fumaric, lactic, mandelic, phthalic, benzenesulfonic,p-tolylsulfonic, citric, tartaric, methanesulfonic, and the like. Alsoincluded are salts of amino acids such as arginate and the like, andsalts of organic acids like glucuronic or galactunoric acids and thelike (see, for example, Berge et al., “Pharmaceutical Salts”, Journal ofPharmaceutical Science 66: 1-19 (1977)). Certain specific compounds ofthe present invention contain both basic and acidic functionalities thatallow the compounds to be converted into either base or acid additionsalts.

The neutral forms of the compounds are preferably regenerated bycontacting the salt with a base or acid and isolating the parentcompounds in the conventional manner. The parent form of the compounddiffers from the various salt forms in certain physical properties, suchas solubility in polar solvents.

In addition to salt forms, the present invention provides compoundswhich are in a prodrug form. Prodrugs of the compounds or complexesdescribed herein readily undergo chemical changes under physiologicalconditions to provide the compounds of the present invention.Additionally, prodrugs can be converted to the compounds of the presentinvention by chemical or biochemical methods in an ex vivo environment.

Certain compounds of the present invention can exist in unsolvated formsas well as solvated forms, including hydrated forms. In general, thesolvated forms are equivalent to unsolvated forms and are encompassedwithin the scope of the present invention. Certain compounds of thepresent invention may exist in multiple crystalline or amorphous forms.In general, all physical forms are equivalent for the uses contemplatedby the present invention and are intended to be within the scope of thepresent invention.

Certain compounds of the present invention possess asymmetric carbonatoms (optical centers) or double bonds; the racemates, diastereomers,geometric isomers and individual isomers are encompassed within thescope of the present invention.

The compounds of the present invention may also contain unnaturalproportions of atomic isotopes at one or more of the atoms thatconstitute such compounds. For example, the compounds may beradiolabeled with radioactive isotopes, such as for example tritium(³H), iodine-125 (¹²⁵I) or carbon-14 (¹⁴C). All isotopic variations ofthe compounds of the present invention, whether radioactive or not, areintended to be encompassed within the scope of the present invention.

The term “pharmaceutically acceptable carrier” or “pharmaceuticallyacceptable vehicle” refers to any formulation or carrier medium thatprovides the appropriate delivery of an effective amount of a activeagent as defined herein, does not interfere with the effectiveness ofthe biological activity of the active agent, and that is sufficientlynon-toxic to the host or patient. Representative carriers include water,oils, both vegetable and mineral, cream bases, lotion bases, ointmentbases and the like. These bases include suspending agents, thickeners,penetration enhancers, and the like. Their formulation is well known tothose in the art of cosmetics and topical pharmaceuticals. Additionalinformation concerning carriers can be found in Remington: The Scienceand Practice of Pharmacy, 21st Ed., Lippincott, Williams & Wilkins(2005) which is incorporated herein by reference.

“Pharmaceutically acceptable topical carrier” and equivalent terms referto pharmaceutically acceptable carriers, as described herein above,suitable for topical application. An inactive liquid or cream vehiclecapable of suspending or dissolving the active agent(s), and having theproperties of being nontoxic and non-inflammatory when applied to theskin, nail, hair, claw or hoof is an example of apharmaceutically-acceptable topical carrier. This term is specificallyintended to encompass carrier materials approved for use in topicalcosmetics as well.

The term “pharmaceutically acceptable additive” refers to preservatives,antioxidants, fragrances, emulsifiers, dyes and excipients known or usedin the field of drug formulation and that do not unduly interfere withthe effectiveness of the biological activity of the active agent, andthat is sufficiently non-toxic to the host or patient. Additives fortopical formulations are well-known in the art, and may be added to thetopical composition, as long as they are pharmaceutically acceptable andnot deleterious to the epithelial cells or their function. Further, theyshould not cause deterioration in the stability of the composition. Forexample, inert fillers, anti-irritants, tackifiers, excipients,fragrances, opacifiers, antioxidants, gelling agents, stabilizers,surfactant, emollients, coloring agents, preservatives, bufferingagents, other permeation enhancers, and other conventional components oftopical or transdermal delivery formulations as are known in the art.

The terms “enhancement,” “penetration enhancement” or “permeationenhancement” relate to an increase in the permeability of the skin,nail, hair, claw or hoof to a drug, so as to increase the rate at whichthe drug permeates through the skin, nail, hair, claw or hoof. Theenhanced permeation effected through the use of such enhancers can beobserved, for example, by measuring the rate of diffusion of the drugthrough animal or human skin, nail, hair, claw or hoof using a diffusioncell apparatus. A diffusion cell is described by Merritt et al.Diffusion Apparatus for Skin Penetration, J of Controlled Release, 1(1984) pp. 161-162. The term “permeation enhancer” or “penetrationenhancer” intends an agent or a mixture of agents, which, alone or incombination, act to increase the permeability of the skin, nail, hair orhoof to a drug.

The term “excipients” is conventionally known to mean carriers, diluentsand/or vehicles used in formulating drug compositions effective for thedesired use.

The term “topical administration” refers to the application of apharmaceutical agent to the external surface of the skin, nail, hair,claw or hoof, such that the agent crosses the external surface of theskin, nail, hair, claw or hoof and enters the underlying tissues.Topical administration includes application of the composition to intactskin, nail, hair, claw or hoof, or to an broken, raw or open wound ofskin, nail, hair, claw or hoof. Topical administration of apharmaceutical agent can result in a limited distribution of the agentto the skin and surrounding tissues or, when the agent is removed fromthe treatment area by the bloodstream, can result in systemicdistribution of the agent.

The term “transdermal delivery” refers to the diffusion of an agentacross the barrier of the skin, nail, hair, claw or hoof resulting fromtopical administration or other application of a composition. Thestratum corneum acts as a barrier and few pharmaceutical agents are ableto penetrate intact skin. In contrast, the epidermis and dermis arepermeable to many solutes and absorption of drugs therefore occurs morereadily through skin, nail, hair, claw or hoof that is abraded orotherwise stripped of the stratum corneum to expose the epidermis.Transdermal delivery includes injection or other delivery through anyportion of the skin, nail, hair, claw or hoof or mucous membrane andabsorption or permeation through the remaining portion. Absorptionthrough intact skin, nail, hair, claw or hoof can be enhanced by placingthe active agent in an appropriate pharmaceutically acceptable vehiclebefore application to the skin, nail, hair, claw or hoof. Passivetopical administration may consist of applying the active agent directlyto the treatment site in combination with emollients or penetrationenhancers. As used herein, transdermal delivery is intended to includedelivery by permeation through or past the integument, i.e. skin, nail,hair, claw or hoof.

II. Introduction

The present invention provides novel boron compounds and methods for thepreparation of these molecules. The invention further provides boroncompounds as analogs comprising a functional moiety, such as a drugmoiety and methods of use for said analogs.

III. The Compounds

In a first aspect, the invention provides a compound having a structureaccording to Formula I:

wherein B is boron. R^(1a) is a member selected from a negative charge,a salt counterion, H, substituted or unsubstituted alkyl, substituted orunsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl,substituted or unsubstituted heterocycloalkyl, substituted orunsubstituted aryl, and substituted or unsubstituted heteroaryl. M1 is amember selected from oxygen, sulfur and NR^(2a). R^(2a) is a memberselected from H, substituted or unsubstituted alkyl, substituted orunsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl,substituted or unsubstituted heterocycloalkyl, substituted orunsubstituted aryl, and substituted or unsubstituted heteroaryl. J1 is amember selected from (CR^(3a)R^(4a))_(n1) and CR^(5a). R^(3a), R^(4a),and R^(5a) are members independently selected from H, OH, NH₂, SH,substituted or unsubstituted alkyl, substituted or unsubstitutedheteroalkyl, substituted or unsubstituted cycloalkyl, substituted orunsubstituted heterocycloalkyl, substituted or unsubstituted aryl, andsubstituted or unsubstituted heteroaryl. The index n1 is an integerselected from 0 to 2. W1 is a member selected from C═O (carbonyl),(CR^(6a)R^(7a))_(m1) and CR^(8a). R^(6a), R^(7a), and R^(8a) are membersindependently selected from H, OH, NH₂, SH, substituted or unsubstitutedalkyl, substituted or unsubstituted heteroalkyl, substituted orunsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl,substituted or unsubstituted aryl, and substituted or unsubstitutedheteroaryl. The index m1 is an integer selected from 0 and 1. A1 is amember selected from CR^(9a) and N. D1 is a member selected fromCR^(10a) and N. E1 is a member selected from CR^(11a) and N. G1 is amember selected from CR^(12a) and N. R^(9a), R^(10a), R^(11a) andR^(12a) are members independently selected from H, OH, NH₂, SH,substituted or unsubstituted alkyl, substituted or unsubstitutedheteroalkyl, substituted or unsubstituted cycloalkyl, substituted orunsubstituted heterocycloalkyl, substituted or unsubstituted aryl, andsubstituted or unsubstituted heteroaryl. The combination of nitrogens(A1+D1+E1+G1) is an integer selected from 0 to 3. A member selected fromR^(3a), R^(4a) and R^(5a) and a member selected from R^(6a), R^(7a) andR^(8a), together with the atoms to which they are attached, areoptionally joined to form a 4 to 7 membered ring. R^(3a) and R^(4a),together with the atoms to which they are attached, are optionallyjoined to form a 4 to 7 membered ring. R^(6a) and R^(7a), together withthe atoms to which they are attached, are optionally joined to form a 4to 7 membered ring. R^(9a) and R^(10a), together with the atoms to whichthey are attached, are optionally joined to form a 4 to 7 membered ring.R^(10a) and R^(11a), together with the atoms to which they are attached,are optionally joined to form a 4 to 7 membered ring. R^(11a) andR^(12a), together with the atoms to which they are attached, areoptionally joined to form a 4 to 7 membered ring. The aspect has theproviso that when M1 is oxygen, W1 is a member selected from(CR^(3a)R^(4a))_(n1), wherein n1 is 0, J1 is a member selected from(CR^(6a)R^(7a))_(m1), wherein m1 is 1, A1 is CR^(9a), D1 is CR^(10a), E1is CR^(11a), G1 is CR^(12a), then R^(9a) is not halogen, methyl, ethyl,or optionally joined with R^(10a) to a form phenyl ring; R^(10a) is notunsubstituted phenoxy, C(CH₃)₃, halogen, CF₃, methoxy, ethoxy, oroptionally joined with R^(9a) to form a phenyl ring; R^(11a) is nothalogen or optionally joined with R^(10a) to form a phenyl ring; andR^(12a) is not halogen. The aspect has the further proviso that when M1is oxygen, W1 is a member selected from (CR^(3a)R^(4a))_(n1), wherein n1is 0, J1 is a member selected from (CR^(6a)R^(7a))_(m1), wherein m1 is1, A1 is CR^(9a), D1 is CR^(10a), E1 is CR^(11a), G1 is CR^(12a), thenneither R^(6a) nor R^(7a) are halophenyl. The aspect has the furtherproviso that when M1 is oxygen, W1 is a member selected from(CR^(3a)R^(4a))_(n1), wherein n1 is 0, J1 is a member selected from(CR^(6a)R^(7a))_(m1), wherein m1 is 1, A1 is CR^(9a), D1 is CR^(10a), E1is CR^(11a), G1 is CR^(12a), and R^(9a), R^(10a) and R^(11a) are H, thenR^(6a), R^(7a) and R^(12a) are not H. The aspect has the further provisothat when M1 is oxygen wherein n1 is 1, J1 is a member selected from(CR^(6a)R^(7a))_(m1), wherein m1 is 0, A1 is CR^(9a), D1 is CR^(10a), E1is CR^(11a), G1 is CR^(12a), R^(9a) is H, R^(10a) is H, R^(11a) is H,R^(6a) is H, R^(7a) is H, R^(12a) is H, then W1 is not C═O (carbonyl).The aspect has the further proviso that when M1 is oxygen, W1 isCR^(5a), J1 is CR^(8a), A1 is CR^(9a), D1 is CR^(10a), E1 is CR^(11a),G1 is CR^(12a), R^(6a), R^(7a), R^(9a), R^(10a), R^(11a) and R^(12a) areH, then R^(5a) and R^(8a), together with the atoms to which they areattached, do not form a phenyl ring.

In an exemplary embodiment, the compound has a structure according toFormula (Ia):

wherein B is boron. R^(1a) is a member selected from a negative charge,a salt counterion, H, substituted or unsubstituted alkyl, substituted orunsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl,substituted or unsubstituted heterocycloalkyl, substituted orunsubstituted aryl, and substituted or unsubstituted heteroaryl. R^(6a)are members independently selected from H, OH, NH₂, SH, substituted orunsubstituted alkyl, substituted or unsubstituted heteroalkyl,substituted or unsubstituted cycloalkyl, substituted or unsubstitutedheterocycloalkyl, substituted or unsubstituted aryl, and substituted orunsubstituted heteroaryl. R^(9a), R^(10a), R^(11a) and R^(12a) aremembers independently selected from H, OH, NH₂, SH, substituted orunsubstituted alkyl, substituted or unsubstituted heteroalkyl,substituted or unsubstituted cycloalkyl, substituted or unsubstitutedheterocycloalkyl, substituted or unsubstituted aryl, and substituted orunsubstituted heteroaryl. R^(9a) and R^(10a), together with the atoms towhich they are attached, are optionally joined to form a 4 to 7 memberedring. R^(10a) and R^(11a), together with the atoms to which they areattached, are optionally joined to form a 4 to 7 membered ring. R^(11a)and R^(12a), together with the atoms to which they are attached, areoptionally joined to form a 4 to 7 membered ring. This embodiment hasthe proviso that R^(9a) is not halogen, methyl, ethyl, or optionallyjoined with R^(10a) to form a 4 to 7 membered ring. This embodiment hasthe proviso that R^(10a) is not unsubstituted phenoxy, C(CH₃)₃, halogen,CF₃, methoxy, ethoxy, optionally joined with R^(9a) to form a 4 to 7membered ring, or optionally joined with R^(11a) to form a 4 to 7membered ring. This embodiment has the proviso that R^(11a) is nothalogen or optionally joined with R^(10a) to form a 4 to 7 memberedring. This embodiment has the proviso that R^(12a) is not halogen.

In an exemplary embodiment, the compound has a structure according toFormula (Ib):

wherein B is boron. R^(x1) is a member selected from substituted orunsubstituted C₁-C₅ alkyl, substituted or unsubstituted C₁-C₅heteroalkyl. R^(y1) and R^(z1) are members independently selected fromH, substituted or unsubstituted alkyl, substituted or unsubstitutedheteroalkyl, substituted or unsubstituted cycloalkyl, substituted orunsubstituted heterocycloalkyl, substituted or unsubstituted aryl, andsubstituted or unsubstituted heteroaryl. R^(6a) are membersindependently selected from H, OH, NH₂, SH, substituted or unsubstitutedalkyl, substituted or unsubstituted heteroalkyl, substituted orunsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl,substituted or unsubstituted aryl, and substituted or unsubstitutedheteroaryl. R^(9a), R^(10a), R^(11a) and R^(12a) are membersindependently selected from H, OH, NH₂, SH, substituted or unsubstitutedalkyl, substituted or unsubstituted heteroalkyl, substituted orunsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl,substituted or unsubstituted aryl, and substituted or unsubstitutedheteroaryl. R^(11a) and R^(12a), together with the atoms to which theyare attached, are optionally joined to form a 4 to 7 membered ring. Thisembodiment has the proviso that when R^(9a), R^(11a) and R^(12a) are H,R^(10a) is not H, halogen, unsubstituted phenoxy or t-butyl. Thisembodiment has the further proviso that when R^(9a) is H, R^(10a) andR^(11a) together with the atoms to which they are attached, are notjoined to form a phenyl ring. This embodiment has the further provisothat when R^(11a) is H, R^(9a) and R^(10a) together with the atoms towhich they are attached, are not joined to form a phenyl ring.

In another aspect, the invention provides a compound having a structureaccording to Formula II:

wherein B is boron. R^(1b) is a member selected from a negative charge,a salt counterion, H, substituted or unsubstituted alkyl, substituted orunsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl,substituted or unsubstituted heterocycloalkyl, substituted orunsubstituted aryl, and substituted or unsubstituted heteroaryl. M2 is amember selected from oxygen, sulfur and NR^(2b). R^(2b) is a memberselected from H, substituted or unsubstituted alkyl, substituted orunsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl,substituted or unsubstituted heterocycloalkyl, substituted orunsubstituted aryl, and substituted or unsubstituted heteroaryl. J2 is amember selected from (CR^(3b)R^(4b))_(n2) and CR^(5b). R^(3b), R^(4b),and R^(5b) are members independently selected from H, OH, NH₂, SH,substituted or unsubstituted alkyl, substituted or unsubstitutedheteroalkyl, substituted or unsubstituted cycloalkyl, substituted orunsubstituted heterocycloalkyl, substituted or unsubstituted aryl, andsubstituted or unsubstituted heteroaryl. The index n2 is an integerselected from 0 to 2. W2 is a member selected from C═O (carbonyl),(CR^(6b)R^(7b))_(m2) and CR^(8b). R^(6b), R^(7b), and R^(8b) are membersindependently selected from H, OH, NH₂, SH, substituted or unsubstitutedalkyl, substituted or unsubstituted heteroalkyl, substituted orunsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl,substituted or unsubstituted aryl, and substituted or unsubstitutedheteroaryl. The index m2 is an integer selected from 0 and 1. A2 is amember selected from CR^(9b) and N. D2 is a member selected fromCR^(10b) and N. E2 is a member selected from CR^(11b) and N. G2 is amember selected from CR^(12b) and N. R^(9b), R^(10b), R^(11b) andR^(12b) are members independently selected from H, OH, NH₂, SH,substituted or unsubstituted alkyl, substituted or unsubstitutedheteroalkyl, substituted or unsubstituted cycloalkyl, substituted orunsubstituted heterocycloalkyl, substituted or unsubstituted aryl, andsubstituted or unsubstituted heteroaryl. The combination of nitrogens(A2+D2+E2+G2) is an integer selected from 0 to 3. A member selected fromR^(3b), R^(4b) and R^(5b) and a member selected from R^(6b), R^(7b) andR^(8b), together with the atoms to which they are attached, areoptionally joined to form a 4 to 7 membered ring. R^(3b) and R^(4b),together with the atoms to which they are attached, are optionallyjoined to form a 4 to 7 membered ring. R^(6b) and R^(7b), together withthe atoms to which they are attached, are optionally joined to form a 4to 7 membered ring. R^(9b) and R^(10b), together with the atoms to whichthey are attached, are optionally joined to form a 4 to 7 membered ring.R^(10b) and R^(11b), together with the atoms to which they are attached,are optionally joined to form a 4 to 7 membered ring. R^(11b) andR^(12b), together with the atoms to which they are attached, areoptionally joined to form a 4 to 7 membered ring.

In an exemplary embodiment, the aspect has the proviso that when M2 isoxygen, W2 is a member selected from (CR^(3b)R^(4b))_(n2), wherein n2 is0, J2 is a member selected from (CR^(6b)R^(7b))_(m2), wherein m2 is 1,A2 is CR^(9b), D2 is CR^(10b), E is CR^(11b), G is CR^(12b), then R^(9b)is not a member selected from halogen, methyl, ethyl, or optionallyjoined with R^(10b) to a form phenyl ring. In another exemplaryembodiment, the aspect has the proviso that when M2 is oxygen, W2 is amember selected from (CR^(3b)R^(4b))_(n), wherein n2 is 0, J2 is amember selected from (CR^(6b)R^(7b))_(m), wherein m2 is 1, A2 isCR^(9b), D2 is CR^(10b), E2 is CR^(11b), G2 is CR^(12b), then R^(10b) isnot a member selected from unsubstituted phenoxy, C(CH₃)₃, halogen, CF₃,methoxy, ethoxy, or optionally joined with R^(9b) to form a phenyl ring.In another exemplary embodiment, the aspect has the proviso that when M2is oxygen, W2 is a member selected from (CR^(3b)R^(4b))_(n), wherein n2is 0, J2 is a member selected from (CR^(6b)R^(7b))_(m2), wherein m2 is1, A2 is CR^(9b), D2 is CR^(10b), E2 is CR^(11b), G2 is CR^(12b), thenR^(11b) is not a member selected from halogen or optionally joined withR^(10b) to form a phenyl ring. In another exemplary embodiment, theaspect has the proviso that when M2 is oxygen, W2 is a member selectedfrom (CR^(3b)R^(4b))_(n2), wherein n2 is 0, J2 is a member selected from(CR^(6b)R^(7b))_(m2), wherein m2 is 1, A2 is CR^(9b), D2 is CR^(10b), E2is CR^(11b), G2 is CR^(12b), then R^(12b) is not halogen. In anotherexemplary embodiment, the aspect has the proviso that when M2 is oxygen,W2 is a member selected from (CR^(3b)R^(4b))_(n2), wherein n2 is 0, J2is a member selected from (CR^(6b)R^(7b))_(m2), wherein m2 is 1, A2 isCR^(9b), D2 is CR^(10b), E2 is CR^(11b), G2 is CR^(12b), then R^(6b) isnot halophenyl. In another exemplary embodiment, the aspect has theproviso that when M2 is oxygen, W2 is a member selected from(CR^(3b)R⁴)_(n2), wherein n2 is 0, J2 is a member selected from(CR^(6b)R^(7b))_(m2), wherein m2 is 1, A2 is CR^(9b), D2 is CR^(10b), E2is CR^(11b), G2 is CR^(12b), then R^(7b) is not halophenyl. In anotherexemplary embodiment, the aspect has the proviso that when M2 is oxygen,W2 is a member selected from (CR^(3b)R^(4b))_(n2), wherein n2 is 0, J2is a member selected from (CR^(6b)R^(7b))_(m2), wherein m2 is 1, A2 isCR^(9b), D2 is C^(10b), E2 is CR^(11b), G2 is CR^(12b), then R^(6b) andR^(7b) are not halophenyl. In another exemplary embodiment, the aspecthas the proviso that when M2 is oxygen, W2 is a member selected from(CR^(3b)R^(4b))_(n2), wherein n2 is 0, J2 is a member selected from(CR^(6b)R^(7b))_(m2), wherein m2 is 1, A2 is CR^(9b), D2 is CR^(10b), E2is CR^(11b), G2 is CR^(12b), and R^(9b), R^(10b) and R^(11b) are H, thenR^(6b), R^(7b) and R^(12b) are not H. In another exemplary embodiment,the aspect has the proviso that when M2 is oxygen wherein n2 is 1, J2 isa member selected from (CR^(6b)R^(7b))_(m2), wherein m2 is 0, A2 isCR^(9b), D2 is CR^(10b), E2 is CR^(11b), G2 is CR^(12b), R^(9b) is H,R^(10b) is H, R^(11b) is H, R^(6b) is H, R^(7b) is H, R^(12b) is H, thenW2 is not C═O (carbonyl). In another exemplary embodiment, the aspecthas the proviso that when M2 is oxygen, W2 is CR^(5b), J2 is CR^(8b), A2is CR^(9b), D2 is CR^(10b), E2 is CR^(11b), G2 is CR^(12b), R^(6b),R^(7b), R^(9b), R^(10b), R^(11b) and R^(12b) are H, then R^(5b) andR^(8b), together with the atoms to which they are attached, do not forma phenyl ring.

In an exemplary embodiment, the compound with a structure according toFormula (IIa):

In another exemplary embodiment, the compound has a structure accordingto Formula (IIb):

wherein R^(7b) is a member selected from H, methyl, ethyl and phenyl.R^(10b) is a member selected from H, OH, NH₂, SH, halogen, substitutedor unsubstituted phenoxy, substituted or unsubstituted phenylalkyloxy,substituted or unsubstituted phenylthio and substituted or unsubstitutedphenylalkylthio. R^(11b) is a member selected from H, OH, NH₂, SH,methyl, substituted or unsubstituted phenoxy, substituted orunsubstituted phenylalkyloxy, substituted or unsubstituted phenylthioand substituted or unsubstituted phenylalkylthio.

In another exemplary embodiment, R^(1b) is a member selected from anegative charge, H and a salt counterion. In another exemplaryembodiment, R^(10b) and R^(11b) are H. In another exemplary embodiment,one member selected from R^(10b) and R^(11b) is H and the other memberselected from R^(10b) and R^(11b) is a member selected from halo,methyl, cyano, methoxy, hydroxymethyl and p-cyanophenyloxy. In anotherexemplary embodiment, R^(10b) and R^(11b) are members independentlyselected from fluoro, chloro, methyl, cyano, methoxy, hydroxymethyl, andp-cyanophenyl. In another exemplary embodiment, R^(1b) is a memberselected from a negative charge, H and a salt counterion; R^(7b) is H;R^(10b) is F and R^(11b) is H. In another exemplary embodiment, R^(11b)and R^(12b), along with the atoms to which they are attached, are joinedto form a phenyl group. In another exemplary embodiment, R^(1b) is amember selected from a negative charge, H and a salt counterion; R^(7b)is H; R^(10b) is 4-cyanophenoxy; and R^(11b) is H.

In another exemplary embodiment, the compound has a structure accordingto Formula (IIc):

wherein R^(10b) is a member selected from H, halogen, CN and substitutedor unsubstituted C₁₋₄ alkyl. In another exemplary embodiment, thecompound has a formulation which is a member selected from:

In another exemplary embodiment, the compound has a structure accordingto Formula (IId):

wherein B is boron. R^(x2) is a member selected from substituted orunsubstituted C₁-C₅ alkyl and substituted or unsubstituted C₁-C₅heteroalkyl. R^(y2) and R^(z2) are members independently selected fromH, substituted or unsubstituted alkyl, substituted or unsubstitutedheteroalkyl, substituted or unsubstituted cycloalkyl, substituted orunsubstituted heterocycloalkyl, substituted or unsubstituted aryl, andsubstituted or unsubstituted heteroaryl.

The compounds of Formulae (I) or (II) can form a hydrate with water,solvates with alcohols such as methanol, ethanol, propanol, and thelike; adducts with amino compounds, such as ammonia, methylamine,ethylamine, and the like; adducts with acids, such as formic acid,acetic acid and the like; complexes with ethanolamine, quinoline, aminoacids, and the like.

Preparation of Boron-containing Small Molecules

The following exemplary schemes illustrate methods of preparingboron-containing molecules of the present invention. These methods arenot limited to producing the compounds shown, but can be used to preparea variety of molecules such as the compounds and complexes describedherein. The compounds of the present invention can also be synthesizedby methods not explicitly illustrated in the schemes but are well withinthe skill of one in the art. The compounds can be prepared using readilyavailable materials of known intermediates.

In the following schemes, the symbol X represents bromo or iodo. Thesymbol Y is selected from H, lower alkyl, and arylalkyl. The symbol Z isselected from H, alkyl, and aryl. The symbol PG represents protectinggroup. The symbols A, D, E, G, R^(x), R^(y), R^(z), R¹, R², R³, R⁴, R⁵,R⁶, R⁷, R⁸, R⁹, R¹⁰, R¹¹, and R¹² can be used to refer to thecorresponding symbols in Formulae (I) or (II). For example, the symbol Acan refer to A1 of Formula (I), or A2 of Formula (II), subject to theprovisos of each Formula.

Preparation Strategy #1

In Scheme 1, Step 1 and 2, compounds 1 or 2 are converted into alcohol3. In step 1, compound 1 is treated with a reducing agent in anappropriate solvent. Suitable reducing agents include borane complexes,such as borane-tetrahydrofuran, borane-dimethylsulfide, combinationsthereof and the like. Lithium aluminum hydride, or sodium borohydridecan also be used as reducing agents. The reducing agents can be used inquantities ranging from 0.5 to 5 equivalents, relative to compound 1 or2. Suitable solvents include diethyl ether, tetrahydrofuran,1,4-dioxane, 1,2-dimethoxyethane, combinations thereof and the like.Reaction temperatures range from 0° C. to the boiling point of thesolvent used; reaction completion times range from 1 to 24 h.

In Step 2, the carbonyl group of compound 2 is treated with a reducingagent in an appropriate solvent. Suitable reducing agents include boranecomplexes, such as borane-tetrahydrofuran, borane-dimethylsulfide,combinations thereof and the like. Lithium aluminum hydride, or sodiumborohydride can also be used as reducing agents. The reducing agents canbe used in quantities ranging from 0.5 to 5 equivalents, relative tocompound 2. Suitable solvents include lower alcohol, such as methanol,ethanol, and propanol, diethyl ether, tetrahydrofuran, 1,4-dioxane and1,2-dimethoxyethane, combinations thereof and the like. Reactiontemperatures range from 0° C. to the boiling point of the solvent used;reaction completion times range from 1 to 24 h.

In Step 3, the hydroxyl group of compound 3 is protected with aprotecting group which is stable under neutral or basic conditions. Theprotecting group is typically selected from methoxymethyl, ethoxyethyl,tetrahydropyran-2-yl, trimethylsilyl, tert-butyldimethylsilyl,tributylsilyl, combinations thereof and the like. In the case ofmethoxymethyl, compound 3 is treated with 1 to 3 equivalents ofchloromethyl methyl ether in the presence of a base. Suitable basesinclude sodium hydride, potassium tert-butoxide, tertiary amines, suchas diisopropylethylamine, triethylamine,1,8-diazabicyclo[5,4,0]undec-7-ene, and inorganic bases, such as sodiumhydroxide, sodium carbonate, potassium hydroxide, potassium carbonate,combinations thereof and the like. The bases can be used in quantitiesranging from 1 to 3 equivalents, relative to compound 3. Reactiontemperatures range from 0° C. to the boiling point of the solvent used;preferably between 0 and 40° C.; reaction completion times range from 1to 48 h.

In the case of tetrahydropyran-2-yl, compound 3 is treated with 1 to 3equivalents of 3,4-dihydro-2H-pyran in the presence of 1 to 10 mol % ofacid catalyst. Suitable acid catalysts include pyridiniump-toluenesulfonic acid, p-toluenesulfonic acid, camphorsulfonic acid,hydrogen chloride, sulfuric acid, combinations thereof and the like.Suitable solvents include dichloromethane, chloroform, tetrahydrofuran,1,4-dioxane, 1,2-dimethoxyethane, toluene, benzene, and acetonitrilecombinations thereof and the like. Reaction temperatures range from 0°C. to the boiling point of the solvent used; preferably between 0 and40° C., and is complete in 1 to 48 h.

In the case of trialkylsilyl, compound 3 is treated with 1 to 3equivalents of chlorotrialkylsilyane in the presence of 1 to 3equivalents of base. Suitable bases include tertiary amines, such asimidazole, diisopropylethylamine, triethylamine,1,8-diazabicyclo[5,4,0]undec-7-ene, combinations thereof and the like.Reaction temperatures range from 0° C. to the boiling point of thesolvent used; preferably between 0 and 40° C.; reaction completion timesrange from 1 to 48 h.

In Step 4, compound 4 is converted into boronic acid (5) through halogenmetal exchange reaction. Compound 4 is treated with 1 to 3 equivalentsof alkylmetal reagent relative to compound 4, such as n-butyllithium,sec-butyllithium, tert-butyllithium, or isopropylmagnesium chloridefollowed by the addition of 1 to 3 equivalents of trialkyl boraterelative to compound 4, such as trimethyl borate, triisopropyl borate,or tributyl borate. Suitable solvents include tetrahydrofuran, ether,1,4-dioxane, 1,2-dimethoxyethane, toluene, hexanes, combinations thereofand the like. Alkylmetal reagent may also be added in the presence oftrialkyl borate. The addition of butyllithium is carried out at between−100 and 0° C., preferably at between −80 and −40° C. The addition ofisopropylmagnesium chloride is carried out at between −80 and 40° C.,preferably at between −20 and 30° C. After the addition of trialkylborate, the reaction is allowed to warm to room temperature, which istypically between 15 and 30° C. When alkylmetal reagent is added in thepresence of trialkyl borate, the reaction mixture is allowed to warm toroom temperature after the addition. Reaction completion times rangefrom 1 to 12 h. Compound 5 may not be isolated and may be used for thenext step without purification or in one pot.

In Step 5, the protecting group of compound 5 is removed under acidicconditions to give compound of Formulae (I) and (II). Suitable acidsinclude acetic acid, trifluoroacetic acid, hydrochloric acid,hydrobromic acid, sulfuric acid, p-toluenesulfonic acid and the like.The acids can be used in quantities ranging from 0.1 to 20 equivalents,relative to compound 5. When the protecting group is trialkylsilyl,basic reagents, such as tetrabutylammonium fluoride, can also be used.Suitable solvents include tetrahydrofuran, 1,4-dioxane,1,2-dimethoxyethane, methanol, ethanol, propanol, acetonitrile, acetone,combination thereof and the like. Reaction temperatures range from 0° C.to the boiling point of the solvent used; preferably between 10 and 40°C.; reaction completion times range from 0.5 to 48 h.

Preparation Strategy #2

In Scheme 2, Step 6, compound 2 is converted into boronic acid (6) via atransition metal catalyzed cross-coupling reaction. Compound 2 istreated with 1 to 3 equivalents of bis(pinacolato)diboron or4,4,5,5-tetramethyl-1,3,2-dioxaborolane in the presence of transitionmetal catalyst, with the use of appropriate ligand and base asnecessary. Suitable transition metal catalysts includepalladium(II)acetate, palladium(II) acetoacetonate,tetrakis(triphenylphosphine)palladium,dichlorobis(triphenylphosphine)palladium,[1,1′-bis(diphenylphosphino)ferrocen]dichloropalladium(II), combinationsthereof and the like. The catalyst can be used in quantities rangingfrom 1 to 5 mol % relative to compound 2. Suitable ligands includetriphenylphosphine, tri(o-tolyl)phosphine, tricyclohexylphosphine,combinations thereof and the like. The ligand can be used in quantitiesranging from 1 to 5 equivalents relative to compound 2. Suitable basesinclude sodium carbonate, potassium carbonate, potassium phenoxide,triethylamine, combinations thereof and the like. The base can be usedin quantities ranging from 1 to 5 equivalents relative to compound 2.Suitable solvents include N,N-dimethylformamide, dimethylsufoxide,tetrahydrofuran, 1,4-dioxane, toluene, combinations thereof and thelike. Reaction temperatures range from 20° C. to the boiling point ofthe solvent used; preferably between 50 and 150° C.; reaction completiontimes range from 1 to 72 h.

Pinacol ester is then oxidatively cleaved to give compound 6. Pinacolester is treated with sodium periodate followed by acid. Sodiumperiodate can be used in quantities ranging from 2 to 5 equivalentsrelative to compound 6. Suitable solvents include tetrahydrofuran,1,4-dioxane, acetonitrile, methanol, ethanol, combinations thereof andthe like. Suitable acids include hydrochloric acid, hydrobromic acid,sulfuric acid combinations thereof and the like. Reaction temperaturesrange from 0° C. to the boiling point of the solvent used; preferablybetween 0 and 50° C.; reaction completion times range from 1 to 72 h.

In Step 7, the carbonyl group of compound 6 is treated with a reducingagent in an appropriate solvent to give a compound of Formulae (I) and(II). Suitable reducing agents include borane complexes, such asborane-tetrahydrofuran, borane-dimethylsulfide, combinations thereof andthe like. Lithium aluminum hydride, or sodium borohydride can also beused as reducing agents. The reducing agents can be used in quantitiesranging from 0.5 to 5 equivalents, relative to compound 6. Suitablesolvents include lower alcohol, such as methanol, ethanol, and propanol,diethyl ether, tetrahydrofuran, 1,4-dioxane and 1,2-dimethoxyethane,combinations thereof and the like. Reaction temperatures range from 0°C. to the boiling point of the solvent used; reaction completion timesrange from 1 to 24 h.

Preparation Strategy #3

In Scheme 3, Step 8, compounds of Formulae (I) and (II) can be preparedin one step from compound 3. Compound 3 is mixed with trialkyl boratethen treated with alkylmetal reagent. Suitable alkylmetal reagentsinclude n-butyllithium, sec-butyllithium, tert-butyllithium combinationsthereof and the like. Suitable trialkyl borates include trimethylborate, triisopropyl borate, tributyl borate, combinations thereof andthe like. The addition of butyllithium is carried out at between −100and 0° C., preferably at between −80 and −40° C. The reaction mixture isallowed to warm to room temperature after the addition. Reactioncompletion times range from 1 to 12 h. The trialkyl borate can be usedin quantities ranging from 1 to 5 equivalents relative to compound 3.The alkylmetal reagent can be used in quantities ranging from 1 to 2equivalents relative to compound 3. Suitable solvents includetetrahydrofuran, ether, 1,4-dioxane, 1,2-dimethoxyethane, toluene,hexanes, combinations thereof and the like. Reaction completion timesrange from 1 to 12 h. Alternatively, a mixture of compound 3 andtrialkyl borate can be refluxed for 1 to 3 h and the alcohol moleculeformed upon the ester exchange can be distilled out before the additionof alkylmetal reagent.

Preparation Strategy #4

In Scheme 4, Step 10, the methyl group of compound 7 is brominated usingN-bromosuccinimide. N-bromosuccinimide can be used in quantities rangingfrom 0.9 to 1.2 equivalents relative to compound 7. Suitable solventsinclude carbon tetrachloride, tetrahydrofuran, 1,4-dioxane,chlorobenzene, combinations thereof and the like. Reaction temperaturesrange from 20° C. to the boiling point of the solvent used; preferablybetween 50 and 150° C.; reaction completion times range from 1 to 12 h.

In Step 11, the bromomethylene group of compound 8 is converted to thebenzyl alcohol 3. Compound 8 is treated with sodium acetate or potassiumacetate. These acetates can be used in quantities ranging from 1 to 10equivalents relative to compound 8. Suitable solvents includetetrahydrofuran, 1,4-dioxane, N,N-dimethylformamide,N,N-dimethylacetamide, N-methylpyrrolidone, dimethylsulfoxide,combinations thereof and the like. Reaction temperatures range from 20°C. to the boiling point of the solvent used; preferably between 50 and100° C.; reaction completion times range from 1 to 12 h. The resultingacetate is hydrolyzed to compound 3 under basic conditions. Suitablebases include sodium hydroxide, lithium hydroxide, potassium hydroxide,combinations thereof and the like. The base can be used in quantitiesranging from 1 to 5 equivalents relative to compound 8. Suitablesolvents include methanol, ethanol, tetrahydrofuran, water, combinationsthereof and the like. Reaction temperatures range from 20° C. to theboiling point of the solvent used; preferably between 50 and 100° C.;reaction completion times range from 1 to 12 h. Alternatively, compound8 can be directly converted into compound 3 under the similar conditionabove.

Steps 3 through 5 convert compound 3 into a compound of Formulae (I) and(II).

Preparation Strategy #5

In Scheme 5, Step 12, compound 2 is treated with(methoxymethyl)triphenylphosphonium chloride or(methoxymethyl)triphenylphosphonium bromide in the presence of basefollowed by acid hydrolysis to give compound 9. Suitable bases includesodium hydride, potassium tert-butoxide, lithium diisopropylamide,butyllithium, lithium hexamethyldisilazane, combinations thereof and thelike. The (methoxymethyl)triphenylphosphonium salt can be used inquantities ranging from 1 to 5 equivalents relative to compound 2. Thebase can be used in quantities ranging from 1 to 5 equivalents relativeto compound 2. Suitable solvents include tetrahydrofuran,1,2-dimethoxyethane, 1,4-dioxane, ether, toluene, hexane,N,N-dimethylformamide, combinations thereof and the like. Reactiontemperatures range from 0° C. to the boiling point of the solvent used;preferably between 0 and 30° C.; reaction completion times range from 1to 12 h. The enolether formed is hydrolyzed under acidic conditions.Suitable acids include hydrochloric acid, hydrobromic acid, sulfuricacid, and the like. Suitable solvents include tetrahydrofuran,1,2-dimethoxyethane, 1,4-dioxane, methanol, ethanol, combination thereofand the like. Reaction temperatures range from 20° C. to the boilingpoint of the solvent used; preferably between 50 and 100° C.; reactioncompletion times range from 1 to 12 h.

Steps 2 through 5 convert compound 9 into a compound of Formulae (I) and(II).

Preparation Strategy #6

In Scheme 6, compound (I) wherein R¹ is H is converted into compound (I)wherein R¹ is alkyl by mixing with the corresponding alcohol, R¹OH. Thesuitable solvents include tetrahydrofuran, 1,2-dimethoxyethane,1,4-dioxane, toluene, combinations thereof and the like. The alcohol(R¹OH) can be used as the solvent as well. Reaction temperatures rangefrom 20° C. to the boiling point of the solvent used; preferably between50 and 100° C.; reaction completion times range from 1 to 12 h.

Preparation Strategy #7

In Scheme 7, compound (Ia) is converted into its aminoalcohol complex(Ib). Compound (Ia) is treated with HOR¹NR^(1a)R^(1b). The aminoalcoholcan be used in quantities ranging from 1 to 10 equivalents relative tocompound (Ia). Suitable solvents include methanol, ethanol, propanol,tetrahydrofuran, acetone, acetonitrile, 1,2-dimethoxyethane,1,4-dioxane, toluene, N,N-dimethylformamide, water, combination thereofand the like. Reaction temperatures range from 20° C. to the boilingpoint of the solvent used; preferably between 50 and 100° C.; reactioncompletion times range from 1 to 24 h.

The compounds of Formulae (I) or (II) can be converted into hydrates andsolvates by methods similar to those described above.

IV. Methods of Inhibiting Microorganism Growth or Killing Microorganisms

In another aspect, the invention provides a method of inhibiting thegrowth of a microorganism, or killing a microorganism, or both,comprising contacting the microorganism with a compound according toFormulae (I) or (II). Microorganisms are members selected from fungi,yeast, viruses, bacteria and parasites. In another exemplary embodiment,the microorganism is inside, or on the surface of an animal. In anexemplary embodiment, the animal is a member selected from human,cattle, deer, reindeer, goat, honey bee, pig, sheep, horse, cow, bull,dog, guinea pig, gerbil, rabbit, cat, camel, yak, elephant, ostrich,otter, chicken, duck, goose, guinea fowl, pigeon, swan, and turkey. Inanother exemplary embodiment, the animal is a human.

In an exemplary embodiment, the microorganism is a member selected froma fungus and a yeast. In another exemplary embodiment, the fungus oryeast is a member selected from Candida species, Trichophyton species,Microsporium species, Aspergillus species, Cryptococcus species,Blastomyces species, Cocciodiodes species, Histoplasma species,Paracoccidiodes species, Phycomycetes species, Malassezia species,Fusarium species, Epidermophyton species, Scytalidium species,Scopulariopsis species, Alternaria species, Penicillium species,Phialophora species, Rhizopus species, Scedosporium species andZygomycetes class. In another exemplary embodiment, the fungus or yeastis a member selected from Aspergillus fumigatus (A. fumigatus),Blastomyces dermatitidis, Candida Albicans (C. albicans, bothfluconazole sensitive and resistant strains), Candida glabrata (C.glabrata), Candida krusei (C. krusei), Cryptococcus neoformans (C.neoformans), Candida parapsilosis (C. parapsilosis), Candida tropicalis(C. tropicalis), Cocciodiodes immitis, Epidermophyton floccosum (E.floccosum), Fusarium solani (F. solani), Histoplasma capsulatum,Malassezia furfur (M. furfur), Malassezia pachydermatis (M.pachydermatis), Malassezia sympodialis (M. sympodialis), Microsporumaudouinii (M. audouinii), Microsporum canis (M. canis), Microsporumgypseum (M. gypseum), Paracoccidiodes brasiliensis and Phycomycetes spp,Trichophyton mentagrophytes (T. mentagrophytes), Trichophyton rubrum (T.rubrum), Trichophyton tonsurans (T. tonsurans). In another exemplaryembodiment, the fungus or yeast is a member selected from Trichophytonconcentricum, T. violaceum, T. schoenleinii, T. verrucosum, T.soudanense, Microsporum gypseum, M. equinum, Candida guilliermondii,Malassezia globosa, M. obtuse, M. restricta, M. slooffiae, andAspergillus flavus. In another exemplary embodiment, the fungus or yeastis a member selected from dermatophytes, Trichophyton, Microsporum,Epidermophyton and yeast-like fungi.

In an exemplary embodiment, the microorganism is a bacteria. In anexemplary embodiment, the bacteria is a gram-positive bacteria. Inanother exemplary embodiment, the gram-positive bacteria is a memberselected from Staphylococcus species, Streptococcus species, Bacillusspecies, Mycobacterium species, Corynebacterium species(Propionibacterium species), Clostridium species, Actinomyces species,Enterococcus species and Streptomyces species. In another exemplaryembodiment, the bacteria is a gram-negative bacteria. In anotherexemplary embodiment, the gram-negative bacteria is a member selectedfrom Acinetobacter species, Neisseria species, Pseudomonas species,Brucella species, Agrobacterium species, Bordetella species, Escherichiaspecies, Shigelia species, Yersinia species, Salmonella species,Klebsiella species, Enterobacter species, Haemophilus species,Pasteurella species, Streptobacillus species, spirochetal species,Campylobacter species, Vibrio species and Helicobacter species. Inanother exemplary embodiment, the bacterium is a member selected fromPropionibacterium acnes; Staphylococcus aureus; Staphylococcusepidermidis, Staphylococcus saprophyticus; Streptococcus pyogenes;Streptococcus agalactiae; Streptococcus pneumoniae; Enterococcusfaecalis; Enterococcus faecium; Bacillus anthracis; Mycobacteriumavium-intracellulare; Mycobacterium tuberculosis, Acinetobacterbaumanii; Corynebacterium diphtheria; Clostridium perfringens;Clostridium botulinum; Clostridium tetani; Neisseria gonorrhoeae;Neisseria meningitidis; Pseudomonas aeruginosa; Legionella pneumophila;Escherichia coli; Yersinia pestis; Haemophilus influenzae; Helicobacterpylori; Campylobacter fetus; Campylobacter jejuni; Vibrio cholerae;Vibrio parahemolyticus; Trepomena pallidum; Actinomyces israelii;Rickettsia prowazekii; Rickettsia rickettsii; Chlamydia trachomatis;Chlamydia psittaci; Brucella abortus; Agrobacterium tumefaciens; andFrancisella tularensis.

In an exemplary embodiment, the microorganism is a bacteria, which is amember selected from acid-fast bacterium, including Mycobacteriumspecies; bacilli, including Bacillus species, Corynebacterium species(also Propionibacterium) and Clostridium species; filamentous bacteria,including Actinomyces species and Streptomyces species; bacilli, such asPseudomonas species, Brucella species, Agrobacterium species, Bordetellaspecies, Escherichia species, Shigella species, Yersinia species,Salmonella species, Klebsiella species, Enterobacter species,Haemophilus species, Pasteurella species, and Streptobacillus species;spirochetal species, Campylobacter species, Vibrio species; andintracellular bacteria including Rickettsiae species and Chlamydiaspecies.

In an exemplary embodiment, the microorganism is a virus. In anexemplary embodiment, the virus is a member selected from hepatitis A-B,human rhinoviruses, Yellow fever virus, human respiratory coronaviruses,Severe acute respiratory syndrome (SARS), respiratory syncytial virus,influenza viruses, parainfluenza viruses 1-4, human immunodeficiencyvirus 1 (HIV-1), human immunodeficiency virus 2 (HIV-2), Herpes simplexvirus 1 (HSV-1), Herpes simplex virus 2 (HSV-2), human cytomegalovirus(HCMV), Varicella zoster virus, Epstein-Barr (EBV), polioviruses,coxsackieviruses, echoviruses, rubella virus, neuroderma-tropic virus,variola virus, papoviruses, rabies virus, dengue virus, West Nile virusand SARS virus. In another exemplary embodiment, the virus is a memberselected from picornaviridae, Flaviviridae, coronaviridae,paramyxoviridae, orthomyxoviridae, retroviridae, herpesviridae andhepadnaviridae. In another exemplary embodiment, the virus is a memberselected from a virus included in the following table:

TABLE A Viruses Virus Category Pertinent Human Infections RNA VirusesPicomaviridae Polio Human hepatitis A Human rhinovirus Togaviridae andRubella - German measles Flaviviridae Yellow fever Coronaviridae Humanrespiratory coronavirus (HCV) Severe acute respiratory syndrome (SAR)Rhabdoviridae Lyssavirus - Rabies Paramyxoviridae Paramyxovirus - MumpsMorbillvirus - measles Pneumovirus - respiratory syncytial virusOrthomyxoviridae Influenza A-C Bunyaviridae Bunyavirus - Bunyamwera(BUN) Hantavirus - Hantaan (HTN) Nairevirus - Crimean-Congo hemorrhagicfever (CCHF) Phlebovirus - Sandfly fever (SFN) Uukuvirus - Uukuniemi(UUK) Rift Valley Fever (RVFN) Arenaviridae Junin - Argentinehemorrhagic fever Machupo - Bolivian hemorrhagic fever Lassa - Lassafever LCM - aseptic lymphocyctic choriomeningitis Reoviridae RotovirusReovirus Orbivirus Retroviridae Human immunodeficiency virus 1 (HIV-1)Human immunodeficiency virus 2 (HIV-2) Simian immunodeficiency virus(SIV) DNA Viruses Papovaviridae Pediatric viruses that reside in kidneyAdenoviridae Human respiratory distress and some deep-seated eyeinfections Parvoviridae Human gastro-intestinal distress (Norwalk Virus)Herpesviridae Herpes simplex virus 1 (HSV-1) Herpes simplex virus 2(HSV-2) Human cytomegalovirus (HCMV) Varicella zoster virus (VZV)Epstein-Barr virus (EBV) Human herpes virus 6 (HHV6) PoxviridaeOrthopoxvirus is sub-genus for smallpox Hepadnaviridae Hepatitis B virus(HBV) Hepatitis C virus (HCV)

In another exemplary embodiment, the microorganism is a parasite. In anexemplary embodiment, the parasite is a member selected from Plasmodiumfalciparum, P. vivax, P. ovale P. malariae, P. berghei, Leishmaniadonovani, L. infantum, L. chagasi, L. mexicana, L. amazonensis, L.venezuelensis, L. tropics, L. major, L. minor, L. aethiopica, L. Bianabraziliensis, L. (V.) guyanensis, L. (V.) panamensis, L. (V.) peruviana,Trypanosoma brucei rhodesiense, T. brucei gambiense, T. cruzi, Giardiaintestinalis, G. lambda, Toxoplasma gondii, Entamoeba histolytica,Trichomonas vaginalis, Pneumocystis carinii, and Cryptosporidium parvum.

V. Methods of Treating or Preventing Infections

In another aspect, the invention provides a method of treating orpreventing an infection, or both. The method includes administering tothe animal a therapeutically effective amount of the compound of theinvention, sufficient to treat or prevent said infection. In anexemplary embodiment, the compound of the invention is according toFormulae (I) or (II). In another exemplary embodiment, the animal is amember selected from human, cattle, deer, reindeer, goat, honey bee,pig, sheep, horse, cow, bull, dog, guinea pig, gerbil, rabbit, cat,camel, yak, elephant, ostrich, otter, chicken, duck, goose, guinea fowl,pigeon, swan, and turkey. In another exemplary embodiment, the animal isa human. In another exemplary embodiment, the animal is a memberselected from a human, cattle, goat, pig, sheep, horse, cow, bull, dog,guinea pig, gerbil, rabbit, cat, chicken and turkey. In anotherexemplary embodiment, the infection is a member selected from a systemicinfection, a cutaneous infection, and an ungual or periungual infection.

V. a) Methods of Treating of Preventing Ungual and/or PeriungualInfections

In another aspect, the invention provides a method of treating orpreventing an ungual and/or periungual infection. The method includesadministering to the animal a therapeutically effective amount of thecompound of the invention, sufficient to treat or prevent saidinfection. In another exemplary embodiment, the method includesadministering the compound of the invention at a site which is a memberselected from the skin, nail, hair, hoof, claw and the skin surroundingthe nail, hair, hoof and claw.

V. a) 1) Onychomycosis

Onychomycosis is a disease of the nail caused by yeast, dermatophytes,or other molds, and represents approximately 50% of all nail disorders.Toenail infection accounts for approximately 80% of onychomycosisincidence, while fingernails are affected in about 20% of the cases.Dermatophytes are the most frequent cause of nail plate invasion,particularly in toenail onychomycosis. Onychomycosis caused by adermatophyte is termed Tinea unguium. Trichophyton rubrum is by far themost frequently isolated dermatophyte, followed by T. mentagrophytes.Distal subungual onychomycosis is the most common presentation of tineaunguium, with the main site of entry through the hyponychium (thethickened epidermis underneath the free distal end of a nail)progressing in time to involve the nail bed and the nail plate.Discoloration, onycholysis, and accumulation of subungual debris andnail plate dystrophy characterize the disease. The disease adverselyaffects the quality of life of its victims, with subject complaintsranging from unsightly nails and discomfort with footwear, to moreserious complications including secondary bacterial infections.

Many methods are known for the treatment of fungal infections, includingthe oral and topical use of antibiotics (e.g., nystatin and amphotericinB), imidazole anti-fungal agents such as miconazole, clotrimazole,fluconazole, econazole and sulconazole, and non-imidazole fungal agentssuch as the allylamine derivatives terbinafine and naftifine, and thebenzylamine butenafine.

However, onychomycosis has proven to be resistant to most treatments.Nail fungal infections reside in an area difficult to access byconventional topical treatment and anti-fungal drugs cannot readilypenetrate the nail plate to reach the infection sites under the nail.Therefore, onychomycosis has traditionally been treated by oraladministration of anti-fungal drugs; however, clearly this isundesirable due to the potential for side effects of such drugs, inparticular those caused by the more potent anti-fungal drugs such asitraconazole and ketoconazole. An alternative method of treatment ofonychomycosis is by removal of the nail before treating with a topicallyactive anti-fungal agent; such a method of treatment is equallyundesirable. Systemic antimycotic agents require prolonged use and havethe potential for significant side effects. Topical agents have usuallybeen of little benefit, primarily because of poor penetration of theanti-fungal agents into and through the nail mass.

In an exemplary embodiment, the invention provides a method of treatingor preventing onychomycosis. The method includes administering to theanimal a therapeutically effective amount of a pharmaceuticalformulation of the invention, sufficient to treat or preventonychomycosis. In another exemplary embodiment, the method includesadministering the pharmaceutical formulation of the invention at a sitewhich is a member selected from the skin, nail, hair, hoof, claw and theskin surrounding the nail, hair, hoof and claw. In another exemplaryembodiment, the pharmaceutical formulation includes a compound having astructure according to Formula (IIb). In another exemplary embodiment,R^(1b) is H. In another exemplary embodiment, R^(10b) and R^(11b) are H.In another exemplary embodiment, one member selected from R^(10b) andR^(11b) is H and the other member selected from R^(10b) and R^(11b) is amember selected from halo, methyl, cyano, methoxy, hydroxymethyl andp-cyanophenyloxy. In another exemplary embodiment, R^(10b) and R^(11b)are members independently selected from fluoro, chloro, methyl, cyano,methoxy, hydroxymethyl, and p-cyanophenyl. In another exemplaryembodiment, R^(1b) is H; R^(7b) is H; R^(10b) is F and R^(11b) are H. Inanother exemplary embodiment, R^(11b) and R^(12b), along with the atomsto which they are attached, are joined to form a phenyl group.

V. a) 2) Other Unugal and Periungual Infections

In an exemplary embodiment, the invention provides a method of treatingor preventing an ungual or periungual infection in a mammal. This methodcomprising administering to the mammal a therapeutically effectiveamount of a compound of the invention, thereby treating or preventingthe ungual or periungual infection. In an exemplary embodiment, theungual or periungual infection is a member selected from: chloronychia,paronychias, erysipeloid, onychorrhexis, gonorrhea, swimming-poolgranuloma, larva migrans, leprosy, Orf nodule, milkers' nodules,herpetic whitlow, acute bacterial perionyxis, chronic perionyxis,sporotrichosis, syphilis, tuberculosis verrucosa cutis, tularemia,tungiasis, peri- and subungual warts, zona, nail dystrophy(trachyonychia), and dermatological diseases with an effect on thenails, such as psoriasis, pustular psoriasis, alopecia aerata,parakeratosis pustulosa, contact dermatosis, Reiter's syndrome,psoriasiform acral dermatitis, lichen planus, idiopathy atrophy in thenails, lichin nitidus, lichen striatus, inflammatory linear verrucousepidermal naevus (ILVEN), alopecia, pemphigus, bullous pemphigoid,acquired epidermolysis bullosa, Darier's disease, pityriasis rubrapilaris, palmoplantar keratoderma, contact eczema, polymorphic erythema,scabies, Bazex syndrome, systemic scleroderma, systemic lupuserythematosus, chronic lupus erythematosus, dermatomyositus.

The compounds and pharmaceutical formulations of the invention usefulfor ungual and periungual applications also find application in thecosmetics field, in particular for the treatment of irregularities ofthe nails, koilonychias, Beau's lines, longitudinal ridging, ingrownnails.

In an exemplary embodiment, the infection is of the skin, nail, hair,claw or hoof, hair, ear and eye and is a member selected fromSporotrichosis, Mycotic keratitis, Extension oculomycosis, Endogenousoculomycosis, Lobomycosis, Mycetoma, Piedra, Pityriasis versicolor,Tinea corporis, Tinea cruris, Tinea pedis, Tinea barbae, Tinea capitis,Tinea nigra, Otomycosis, Tinea favosa, Chromomycosis, and TineaImbricata.

V. b) Methods of Treating Systemic Diseases

In another aspect, the invention provides a method of treating asystemic disease. The method involves contacting an animal with acompound of the invention. The method of delivery for treatment ofsystemic diseases can be oral, intravenous or transdermal.

In an exemplary embodiment, the infection is systemic and is a memberselected from candidiasis, aspergillosis, coccidioidomycosis,cryptococcosis, histoplasmosis, blastomycosis, paracoccidioidomycosis,zygomycosis, phaeohyphomycosis and rhinosporidiosis.

V. c) Methods of Treating Diseases Involving Viruses

The compounds of the invention are useful for the treatment of diseasesof both animals and humans, involving viruses. In an exemplaryembodiment, the disease is a member selected from hepatitis A—B—C,yellow fever, respiratory syncytial, influenza, AIDS, herpes simplex,chicken pox, varicella zoster, and Epstein-Barr disease.

V. d) Methods of Treating Diseases Involving Parasites

The compounds of the invention are useful for the treatment of diseasesof both animals and humans, involving parasites. In an exemplaryembodiment, the disease is a member selected from malaria, Chagas'disease, Leishmaniasis, African sleeping sickness (African humantrypanosomiasis), giardiasis, toxoplasmosis, amebiasis andcryptosporidiosis.

VI. Methods of Nail Penetration

It is believed that poor penetration of the active agent through thehoof or nail plate and/or excessive binding to keratin, (the majorprotein in nails and hair) are the reasons for the poor efficacy of 8%ciclopirox w/w in commercial lacquer and other topical treatments thathave failed in clinical trials. In mild cases of onychomycosis, thepathogenic fungi reside in the nail plate only. In moderate to severecases the pathogenic fungi establish a presence in the nail plate and inthe nail bed. If the infection is cleared from the nail plate but notfrom the nail bed, the fungal pathogen can re-infect the nail plate.Therefore, to effectively treat onychomycosis, the infection must beeliminated from the nail plate and the nail bed. To do this, the activeagent must penetrate and disseminate substantially throughout the nailplate and nail bed.

It is believed that in order for an active agent to be effective oncedisseminated throughout the infected area, it must be bioavailable tothe fungal pathogen and cannot be so tightly and/or preferentially boundto keratin that the drug is rendered inactive.

An understanding of the morphology of the nail plate suggests certainphysicochemical properties of an active agent that would facilitatepenetration of the nail plate. The desired physicochemical propertiesare described throughout. The tested compounds of the present inventionare able to penetrate the nail plate and were also active againstTrichophyton rubrum and mentagrophytes and other species. In addition,the tested compounds are also active against Trichophyton rubrum in thepresence of 5% keratin powder.

In another aspect, the invention provides a method of delivering acompound from the dorsal layer of the nail plate to the nail bed. Thismethod comprises contacting the cell with a compound capable ofpenetrating the nail plate, under conditions sufficient to penetrate thenail. The compound has a molecular weight of between about 100 and about200 Da. The compound also has a log P value of between about 1.0 andabout 2.6. The compound additionally has a water solubility betweenabout 0.1 mg/mL and 1 g/mL octanol/saturated water, thereby deliveringsaid compound.

In a preferred embodiment, the physicochemical properties of thecompound of the invention, described by quantities predictive formigration of the compound through the nail plate, including, but notlimited to, molecular weight, log P and solubility in water, and thelike, are effective to provide substantial penetration of the nailplate.

Compounds with a molecular weight of less than 200 Da penetrate the nailplate in a manner superior to the commercially available treatment foronychomycosis. In one embodiment of the present invention the compoundhas a molecular weight of between 130 and 200. In another embodiment ofthis invention, the compound has a molecular weight of from about 140 toabout 200 Da. In another embodiment of this invention, the compound hasa molecular weight of from about 170 to about 200 Da. In anotherembodiment of this invention, the compound has a molecular weight offrom about 155 to about 190 Da. In another embodiment of this invention,the compound has a molecular weight of from about 165 to about 185 Da.In another embodiment of this invention, the compound has a molecularweight of from about 145 to about 170 Da. In yet another embodiment themolecular weight is either 151.93 or 168.39 Da.

In one embodiment of the present invention the compound has a Log Pvalue of between about −3.5 to about 2.5. In another exemplaryembodiment, the compound has a Log P value of from about −1.0 to about2.5. In another exemplary embodiment, the compound has a Log P value offrom about −1.0 to about 2.0. In another exemplary embodiment, thecompound has a Log P value of from about −0.5 to about 2.5. In anotherexemplary embodiment, the compound has a Log P value of from about −0.5to about 1.5. In another exemplary embodiment, the compound has a Log Pvalue of from about 0.5 to about 2.5. In another exemplary embodiment,the compound has a Log P value of from about 1.0 to about 2.5. In yetanother exemplary embodiment, the compound has a Log P value of 1.9 or2.3.

Also contemplated by the present invention is a compound with a Log Pvalue less then 2.5, with a molecular weight less than 200 Da, that arestill able to penetrate the nail plate.

In one embodiment of the present invention the compound has a watersolubility between about 0.1 mg/mL to 1 g/mL in octanol saturated water.In one embodiment of the present invention the compound has a watersolubility of between 0.1 mg/mL and 100 mg/mL. In another embodiment ofthis invention, the compound has a water solubility of from about 0.1mg/mL and 10 mg/mL. In another embodiment of this invention, thecompound has a water solubility of from about 0.1 mg/mL and 1 mg/mL. Inanother embodiment of this invention, the compound has a watersolubility of from about 5 mg/mL and 1 g/mL. In another embodiment ofthis invention, the compound has a water solubility of from about 10mg/mL and 500 g/mL. In another embodiment of this invention, thecompound has a water solubility of from about 80 mg/mL and 250 mg/mL.

In an exemplary embodiment, the present invention provides a compoundwith a Log P value selected from a range above, with a molecular weightselected from a range above, that are still able to penetrate the nailplate.

In an exemplary embodiment, the present invention provides compoundswith a molecular weight selected from a range above, with a watersolubility selected from a range above, that are still able to penetratethe nail plate.

In an exemplary embodiment, the present invention provides compoundswith a log P selected from a range above, with a water solubilityselected from a range above, that are still able to penetrate the nailplate.

In an exemplary embodiment, the present invention provides compoundswith a molecular weight selected from a range above, with a log Pselected from a range above, and with a water solubility selected from arange above, that are still able to penetrate the nail plate.

Penetration of the nail by the active ingredient may be effected by thepolarity of the formulation. However, the polarity of the formulation isnot expected have as much influence on nail penetration as some of theother factors, such as the molecular weight or the Log P of the activeingredient. The presence of penetration enhancing agents in theformulation is likely to increase penetration of the active agent whencompared to similar formulations containing no penetration enhancingagent

Some examples of molecules with optimal physicochemical properties aregiven in the table below.

Structure:

Formula: C₇H₆BFO₂ C₇H₆BClO₂ Molecular 151.93 168.39 weight (Da): Plasmaprotein 66 83 binding (%): LogP: 1.9 2.3 Water solubility >100 >100(μg/mL):

Compound 3 below is an example of a compound similar in molecular weightto ciclopirox, and like ciclopirox, penetrates the nail plate poorly.

Structure:

Formula: C₁₃H₁₀BFO Molecular weight (Da): 212.03 Plasma protein binding(%): 100 cLogP: 3.55 Water solubility (μg/mL): not determined

In a preferred embodiment the topical formulations including a compoundof Formulae (I) or (II) described structurally above has a totalmolecular weight of less than 200 Da, has a Log P of less than 2.5, anda minimum inhibitory concentration against Trichophyton rubrum that issubstantially unchanged in the presence of 5% keratin.

This invention is still further directed to methods for treating a viralinfection mediated at least in part by dermatophytes, Trichophyton,Microsporum or Epidermophyton species, or a yeast-like fungi includingCandida species, in mammals, which methods comprise administering to amammal, that has been diagnosed with said viral infection or is at riskof developing said viral infection, a pharmaceutical compositioncomprising a pharmaceutically acceptable diluent and a therapeuticallyeffective amount of a compound described herein or mixtures of one ormore of such compounds. In one embodiment the infection isonychomycosis.

Compounds contemplated by the present invention may have broad spectrumantifungal activity and as such may be candidates for use against othercutaneous fungal infections.

The methods provided in this aspect of the invention are useful in thepenetration of nails and hoofs, as well as the treatment of ungual andperiungual conditions.

VII. Pharmaceutical Formulations

In another aspect, the invention is a pharmaceutical formulation whichincludes: (a) a pharmaceutically acceptable excipient; and (b) acompound of the invention. In another aspect, the invention is apharmaceutical formulation which includes: (a) a pharmaceuticallyacceptable excipient; and (b) a compound having a structure according toFormula (I), (Ia), (Ib), (Ic), or (Id). In another aspect, the inventionis a pharmaceutical formulation which includes: (a) a pharmaceuticallyacceptable excipient; and (b) a compound which has a structure accordingto Formula (II), (IIa), (IIb), (IIc), (IId).

In another aspect, the invention is a pharmaceutical formulationcomprising: (a) a pharmaceutically acceptable excipient; and (b) acompound having a structure according to Formula II:

wherein B is boron. R^(1b) is a member selected from a negative charge,a salt counterion, H, substituted or unsubstituted alkyl, substituted orunsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl,substituted or unsubstituted heterocycloalkyl, substituted orunsubstituted aryl, and substituted or unsubstituted heteroaryl. M2 is amember selected from oxygen, sulfur and NR^(2b). R^(2b) is a memberselected from H, substituted or unsubstituted alkyl, substituted orunsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl,substituted or unsubstituted heterocycloalkyl, substituted orunsubstituted aryl, and substituted or unsubstituted heteroaryl. J2 is amember selected from (CR^(3b)R^(4b))_(n2) and CR^(5b). R^(3b), R^(4b),and R^(5b) are members independently selected from H, OH, NH₂, SH,substituted or unsubstituted alkyl, substituted or unsubstitutedheteroalkyl, substituted or unsubstituted cycloalkyl, substituted orunsubstituted heterocycloalkyl, substituted or unsubstituted aryl, andsubstituted or unsubstituted heteroaryl. The index n2 is an integerselected from 0 to 2. W2 is a member selected from C═O (carbonyl),(CR^(6b)R^(7b))_(m2) and CR^(8b). R^(6b), R^(7b), and R^(8b) are membersindependently selected from H, OH, NH₂, SH, substituted or unsubstitutedalkyl, substituted or unsubstituted heteroalkyl, substituted orunsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl,substituted or unsubstituted aryl, and substituted or unsubstitutedheteroaryl. The index m2 is an integer selected from 0 and 1. A2 is amember selected from CR^(9b) and N. D2 is a member selected fromCR^(10b) and N. E2 is a member selected from CR^(11b) and N. G2 is amember selected from CR^(12b) and N. R^(9b), R^(10b), R^(11b) andR^(12b) are members independently selected from H, OH, NH₂, SH,substituted or unsubstituted alkyl, substituted or unsubstitutedheteroalkyl, substituted or unsubstituted cycloalkyl, substituted orunsubstituted heterocycloalkyl, substituted or unsubstituted aryl, andsubstituted or unsubstituted heteroaryl. The combination of nitrogens(A2+D2+E2+G2) is an integer selected from 0 to 3. A member selected fromR^(3b), R^(4b) and R^(5b) and a member selected from R^(6b), R^(7b) andR^(8b), together with the atoms to which they are attached, areoptionally joined to form a 4 to 7 membered ring. R^(3b) and R^(4b),together with the atoms to which they are attached, are optionallyjoined to form a 4 to 7 membered ring. R^(6b) and R^(7b), together withthe atoms to which they are attached, are optionally joined to form a 4to 7 membered ring. R^(9b) and R^(10b), together with the atoms to whichthey are attached, are optionally joined to form a 4 to 7 membered ring.R^(10b) and R^(11b), together with the atoms to which they are attached,are optionally joined to form a 4 to 7 membered ring. R^(11b) andR^(12b), together with the atoms to which they are attached, areoptionally joined to form a 4 to 7 membered ring.

In an exemplary embodiment, the aspect has the proviso that when M2 isoxygen, W2 is a member selected from (CR^(3b)R^(4b))_(n2), wherein n2 is0, J2 is a member selected from (CR^(6b)R^(7b))_(m2), wherein m2 is 1,A2 is CR^(9b), D2 is CR^(10b), E is CR^(11b), G is CR^(12b), then R^(9b)is not a member selected from halogen, methyl, ethyl, or optionallyjoined with R^(10b) to a form phenyl ring. In another exemplaryembodiment, the aspect has the proviso that when M2 is oxygen, W2 is amember selected from (CR^(3b)R^(4b))_(n), wherein n2 is 0, J2 is amember selected from (CR^(6b)R^(7b))_(m), wherein m2 is 1, A2 isCR^(9b), D2 is CR^(10b), E2 is CR^(11b), G2 is CR^(12b), then R^(10b) isnot a member selected from unsubstituted phenoxy, C(CH₃)₃, halogen, CF₃,methoxy, ethoxy, or optionally joined with R^(9b) to form a phenyl ring.In another exemplary embodiment, the aspect has the proviso that when M2is oxygen, W2 is a member selected from (CR^(3b)R^(4b))_(n), wherein n2is 0, J2 is a member selected from (CR^(6b)R^(7b))_(m2), wherein m2 is1, A2 is CR^(9b), D2 is CR^(10b), E2 is CR^(11b), G2 is CR^(12b), thenR^(11b) is not a member selected from halogen or optionally joined withR^(10b) to form a phenyl ring. In another exemplary embodiment, theaspect has the proviso that when M2 is oxygen, W2 is a member selectedfrom (CR^(3b)R^(4b))_(n2), wherein n2 is 0, J2 is a member selected from(CR^(6b)R^(7b))_(m2), wherein m2 is 1, A2 is CR^(9b), D2 is CR^(10b), E2is CR^(11b), G2 is CR^(12b), then R^(12b) is not halogen. In anotherexemplary embodiment, the aspect has the proviso that when M2 is oxygen,W2 is a member selected from (CR^(3b)R^(4b))_(n2), wherein n2 is 0, J2is a member selected from (CR^(6b)R^(7b))_(m2), wherein m2 is 1, A2 isCR^(9b), D2 is CR^(10b), E2 is CR^(11b), G2 is CR^(12b), then R^(6b) isnot halophenyl. In another exemplary embodiment, the aspect has theproviso that when M2 is oxygen, W2 is a member selected from(CR^(3b)R^(4b))_(n2), wherein n2 is 0, J2 is a member selected from(CR^(6b)R^(7b))_(m2), wherein m2 is 1, A2 is CR^(9b), D2 is CR^(10b), E2is CR^(11b), G2 is CR^(12b), then R^(7b) is not halophenyl. In anotherexemplary embodiment, the aspect has the proviso that when M2 is oxygen,W2 is a member selected from (CR^(3b)R^(4b))_(n2), wherein n2 is 0, J2is a member selected from (CR^(6b)R^(7b))_(m2), wherein m2 is 1, A2 isCR^(9b), D2 is CR^(10b), E2 is CR^(11b), G2 is CR^(12b), then R^(6b) andR^(7b) are not halophenyl. In another exemplary embodiment, the aspecthas the proviso that when M2 is oxygen, W2 is a member selected from(CR^(3b)R^(4b))_(n2), wherein n2 is 0, J2 is a member selected from(CR^(6b)R^(7b))_(m2), wherein m2 is 1, A2 is CR^(9b), D2 is CR^(10b), E2is CR^(11b), G2 is CR^(12b), and R^(9b), R^(10b) and R^(11b) are H, thenR^(6b), R^(7b) and R^(12b) are not H. In another exemplary embodiment,the aspect has the proviso that when M2 is oxygen wherein n2 is 1, J2 isa member selected from (CR^(6b)R^(7b))_(m2), wherein m2 is 0, A2 isCR^(9b), D2 is CR^(10b), E2 is CR^(11b), G2 is CR^(12b), R^(9b) is H,R^(10b) is H, R^(11b) is H, R^(6b) is H, R^(7b) is H, R^(12b) is H, thenW2 is not C═O (carbonyl). In another exemplary embodiment, the aspecthas the proviso that when M2 is oxygen, W2 is CR^(5b), J2 is CR^(8b), A2is CR^(9b), D2 is CR^(10b), E2 is CR^(11b), G2 is CR^(12b), R^(6b),R^(7b), R^(9b), R^(10b), R^(11b) and R^(12b) are H, then R^(5b) andR^(8b), together with the atoms to which they are attached, do not forma phenyl ring.

In an exemplary embodiment, the pharmaceutical formulation has acompound with a structure according to Formula (IIa):

In another exemplary embodiment, the pharmaceutical formulation has acompound with a structure according to Formula (IIb):

wherein R^(7b) is a member selected from H, methyl, ethyl and phenyl.R^(10b) is a member selected from H, OH, NH₂, SH, halogen, substitutedor unsubstituted phenoxy, substituted or unsubstituted phenylalkyloxy,substituted or unsubstituted phenylthio and substituted or unsubstitutedphenylalkylthio. R^(11b) is a member selected from H, OH, NH₂, SH,methyl, substituted or unsubstituted phenoxy, substituted orunsubstituted phenylalkyloxy, substituted or unsubstituted phenylthioand substituted or unsubstituted phenylalkylthio.

In another exemplary embodiment, R^(1b) is a member selected from anegative charge, H and a salt counterion. In another exemplaryembodiment, R^(10b) and R^(11b) are H. In another exemplary embodiment,one member selected from R^(10b) and R^(11b) is H and the other memberselected from R^(10b) and R^(11b) is a member selected from halo,methyl, cyano, methoxy, hydroxymethyl and p-cyanophenyloxy. In anotherexemplary embodiment, R^(10b) and R^(11b) are members independentlyselected from fluoro, chloro, methyl, cyano, methoxy, hydroxymethyl, andp-cyanophenyl. In another exemplary embodiment, R^(1b) is a memberselected from a negative charge, H and a salt counterion; R^(7b) is H;R^(10b) is F and R^(11b) is H. In another exemplary embodiment, R^(11b)and R^(12b), along with the atoms to which they are attached, are joinedto form a phenyl group. In another exemplary embodiment, R^(1b) is amember selected from a negative charge, H and a salt counterion; R^(7b)is H; R^(10b) is 4-cyanophenoxy; and R^(11b) is H.

In another exemplary embodiment, the pharmaceutical formulation has acompound with a structure according to Formula (IIc):

wherein R^(10b) is a member selected from H, halogen, CN and substitutedor unsubstituted C₁₋₄ alkyl. In another exemplary embodiment, thecompound has a formulation which is a member selected from:

In another exemplary embodiment, the pharmaceutical formulation has acompound with a structure according to Formula (IId):

wherein B is boron. R^(x2) is a member selected from substituted orunsubstituted C₁-C₅ alkyl and substituted or unsubstituted C₁-C₅heteroalkyl. R² and R^(z2) are members independently selected from H,substituted or unsubstituted alkyl, substituted or unsubstitutedheteroalkyl, substituted or unsubstituted cycloalkyl, substituted orunsubstituted heterocycloalkyl, substituted or unsubstituted aryl, andsubstituted or unsubstituted heteroaryl.

The pharmaceutical formulations of the invention can take a variety offorms adapted to the chosen route of administration. Those skilled inthe art will recognize various synthetic methodologies that may beemployed to prepare non-toxic pharmaceutical formulations incorporatingthe compounds described herein. Those skilled in the art will recognizea wide variety of non-toxic pharmaceutically acceptable solvents thatmay be used to prepare solvates of the compounds of the invention, suchas water, ethanol, propylene glycol, mineral oil, vegetable oil anddimethylsulfoxide (DMSO).

The compositions of the invention may be administered orally, topically,parenterally, by inhalation or spray or rectally in dosage unitformulations containing conventional non-toxic pharmaceuticallyacceptable carriers, adjuvants and vehicles. It is further understoodthat the best method of administration may be a combination of methods.Oral administration in the form of a pill, capsule, elixir, syrup,lozenge, troche, or the like is particularly preferred. The termparenteral as used herein includes subcutaneous injections, intradermal,intravascular (e.g., intravenous), intramuscular, spinal, intrathecalinjection or like injection or infusion techniques.

The pharmaceutical formulations containing compounds of the inventionare preferably in a form suitable for oral use, for example, as tablets,troches, lozenges, aqueous or oily suspensions, dispersible powders orgranules, emulsion, hard or soft capsules, or syrups or elixirs.

Compositions intended for oral use may be prepared according to anymethod known in the art for the manufacture of pharmaceuticalformulations, and such compositions may contain one or more agentsselected from the group consisting of sweetening agents, flavoringagents, coloring agents and preserving agents in order to providepharmaceutically elegant and palatable preparations. Tablets may containthe active ingredient in admixture with non-toxic pharmaceuticallyacceptable excipients that are suitable for the manufacture of tablets.These excipients may be for example, inert diluents, such as calciumcarbonate, sodium carbonate, lactose, calcium phosphate or sodiumphosphate; granulating and disintegrating agents, for example, cornstarch, or alginic acid; binding agents, for example starch, gelatin oracacia; and lubricating agents, for example magnesium stearate, stearicacid or talc. The tablets may be uncoated or they may be coated by knowntechniques to delay disintegration and absorption in thegastrointestinal tract and thereby provide a sustained action over alonger period. For example, a time delay material such as glycerylmonostearate or glyceryl distearate may be employed.

Formulations for oral use may also be presented as hard gelatin capsuleswherein the active ingredient is mixed with an inert solid diluent, forexample, calcium carbonate, calcium phosphate or kaolin, or as softgelatin capsules wherein the active ingredient is mixed with water or anoil medium, for example peanut oil, liquid paraffin or olive oil.

Aqueous suspensions contain the active materials in admixture withexcipients suitable for the manufacture of aqueous suspensions. Suchexcipients are suspending agents, for example sodiumcarboxymethylcellulose, methylcellulose, hydroxypropylmethylcellulose,sodium alginate, polyvinylpyrrolidone, gum tragacanth and gum acacia;and dispersing or wetting agents, which may be a naturally-occurringphosphatide, for example, lecithin, or condensation products of analkylene oxide with fatty acids, for example polyoxyethylene stearate,or condensation products of ethylene oxide with long chain aliphaticalcohols, for example heptadecaethyleneoxycetanol, or condensationproducts of ethylene oxide with partial esters derived from fatty acidsand a hexitol such as polyoxyethylene sorbitol monooleate, orcondensation products of ethylene oxide with partial esters derived fromfatty acids and hexitol anhydrides, for example polyethylene sorbitanmonooleate. The aqueous suspensions may also contain one or morepreservatives, for example ethyl, or n-propyl p-hydroxybenzoate, one ormore coloring agents, one or more flavoring agents, and one or moresweetening agents, such as sucrose or saccharin.

Oily suspensions may be formulated by suspending the active ingredientsin a vegetable oil, for example arachis oil, olive oil, sesame oil orcoconut oil, or in a mineral oil such as liquid paraffin. The oilysuspensions may contain a thickening agent, for example beeswax, hardparaffin or cetyl alcohol. Sweetening agents such as those set forthabove, and flavoring agents may be added to provide palatable oralpreparations. These compositions may be preserved by the addition of ananti-oxidant such as ascorbic acid.

Dispersible powders and granules suitable for preparation of an aqueoussuspension by the addition of water provide the active ingredient inadmixture with a dispersing or wetting agent, suspending agent and oneor more preservatives. Suitable dispersing or wetting agents andsuspending agents are exemplified by those already mentioned above.Additional excipients, for example sweetening, flavoring and coloringagents, may also be present.

Pharmaceutical formulations of the invention may also be in the form ofoil-in-water emulsions and water-in-oil emulsions. The oily phase may bea vegetable oil, for example olive oil or arachis oil, or a mineral oil,for example liquid paraffin or mixtures of these. Suitable emulsifyingagents may be naturally-occurring gums, for example gum acacia or gumtragacanth; naturally-occurring phosphatides, for example soy bean,lecithin, and esters or partial esters derived from fatty acids andhexitol; anhydrides, for example sorbitan monooleate; and condensationproducts of the said partial esters with ethylene oxide, for examplepolyoxyethylene sorbitan monooleate. The emulsions may also containsweetening and flavoring agents.

Syrups and elixirs may be formulated with sweetening agents, for exampleglycerol, propylene glycol, sorbitol or sucrose. Such formulations mayalso contain a demulcent, a preservative, and flavoring and coloringagents. The pharmaceutical formulations may be in the form of a sterileinjectable aqueous or oleaginous suspension. This suspension may beformulated according to the known art using those suitable dispersing orwetting agents and suspending agents, which have been mentioned above.The sterile injectable preparation may also be a sterile injectablesolution or suspension in a non-toxic parenterally acceptable diluent orsolvent, for example as a solution in 1,3-butanediol. Among theacceptable vehicles and solvents that may be employed are water,Ringer's solution and isotonic sodium chloride solution. In addition,sterile, fixed oils are conventionally employed as a solvent orsuspending medium. For this purpose any bland fixed oil may be employedincluding synthetic mono- or diglycerides. In addition, fatty acids suchas oleic acid find use in the preparation of injectables.

The composition of the invention may also be administered in the form ofsuppositories, e.g., for rectal administration of the drug. Thesecompositions can be prepared by mixing the drug with a suitablenon-irritating excipient that is solid at ordinary temperatures butliquid at the rectal temperature and will therefore melt in the rectumto release the drug. Such materials are cocoa butter and polyethyleneglycols.

Alternatively, the compositions can be administered parenterally in asterile medium. The drug, depending on the vehicle and concentrationused, can either be suspended or dissolved in the vehicle.Advantageously, adjuvants such as local anesthetics, preservatives andbuffering agents can be dissolved in the vehicle.

For administration to non-human animals, the composition containing thetherapeutic compound may be added to the animal's feed or drinkingwater. Also, it will be convenient to formulate animal feed and drinkingwater products so that the animal takes in an appropriate quantity ofthe compound in its diet. It will further be convenient to present thecompound in a composition as a premix for addition to the feed ordrinking water. The composition can also added as a food or drinksupplement for humans.

Dosage levels of the order of from about 5 mg to about 250 mg perkilogram of body weight per day and more preferably from about 25 mg toabout 150 mg per kilogram of body weight per day, are useful in thetreatment of the above-indicated conditions. The amount of activeingredient that may be combined with the carrier materials to produce asingle dosage form will vary depending upon the condition being treatedand the particular mode of administration. Dosage unit forms willgenerally contain between from about 1 mg to about 500 mg of an activeingredient.

Frequency of dosage may also vary depending on the compound used and theparticular disease treated. However, for treatment of most disorders, adosage regimen of 4 times daily or less is preferred. It will beunderstood, however, that the specific dose level for any particularpatient will depend upon a variety of factors including the activity ofthe specific compound employed, the age, body weight, general health,sex, diet, time of administration, route of administration and rate ofexcretion, drug combination and the severity of the particular diseaseundergoing therapy.

Preferred compounds of the invention will have desirable pharmacologicalproperties that include, but are not limited to, oral bioavailability,low toxicity, low serum protein binding and desirable in vitro and invivo half-lives. Penetration of the blood brain barrier for compoundsused to treat CNS disorders is necessary, while low brain levels ofcompounds used to treat peripheral disorders are often preferred.

Assays may be used to predict these desirable pharmacologicalproperties. Assays used to predict bioavailability include transportacross human intestinal cell monolayers, including Caco-2 cellmonolayers. Toxicity to cultured hepatocyctes may be used to predictcompound toxicity. Penetration of the blood brain barrier of a compoundin humans may be predicted from the brain levels of laboratory animalsthat receive the compound intravenously.

Serum protein binding may be predicted from albumin binding assays. Suchassays are described in a review by Oravcova, et al. (Journal ofChromatography B (1996) volume 677, pages 1-27).

Compound half-life is inversely proportional to the frequency of dosageof a compound. In vitro half-lives of compounds may be predicted fromassays of microsomal half-life as described by Kuhnz and Gieschen (DrugMetabolism and Disposition, (1998) volume 26, pages 1120-1127).

The amount of the composition required for use in treatment will varynot only with the particular compound selected but also with the routeof administration, the nature of the condition being treated and the ageand condition of the patient and will ultimately be at the discretion ofthe attendant physician or clinician.

VII. a) Topical Formulations

In a preferred embodiment, the methods of the invention can be usedemployed through the topical application of the compounds describedherein.

The compositions of the present invention comprises fluid or semi-solidvehicles that may include but are not limited to polymers, thickeners,buffers, neutralizers, chelating agents, preservatives, surfactants oremulsifiers, antioxidants, waxes or oils, emollients, sunscreens, and asolvent or mixed solvent system. The solvent or mixed solvent system isimportant to the formation because it is primarily responsible fordissolving the drug. The best solvent or mixed solvent systems are alsocapable of maintaining clinically relevant levels of the drug insolution despite the addition of a poor solvent to the formulation. Thetopical compositions useful in the subject invention can be made into awide variety of product types. These include, but are not limited to,lotions, creams, gels, sticks, sprays, ointments, pastes, foams,mousses, and cleansers. These product types can comprise several typesof carrier systems including, but not limited to particles,nanoparticles, and liposomes. If desired, disintegrating agents can beadded, such as the cross-linked polyvinyl pyrrolidone, agar or alginicacid or a salt thereof such as sodium alginate. Techniques forformulation and administration can be found in Remington: The Scienceand Practice of Pharmacy, supra. The formulation can be selected tomaximize delivery to a desired target site in the body.

Lotions, which are preparations that are to be applied to the skin,nail, hair, claw or hoof surface without friction, are typically liquidor semi-liquid preparations in which finely divided solid, waxy, orliquid are dispersed. Lotions will typically contain suspending agentsto produce better dispersions as well as compounds useful for localizingand holding the active agent in contact with the skin, nail, hair, clawor hoof, e.g., methylcellulose, sodium carboxymethyl-cellulose, or thelike.

Creams containing the active agent for delivery according to the presentinvention are viscous liquid or semisolid emulsions, either oil-in-wateror water-in-oil. Cream bases are water-washable, and contain an oilphase, an emulsifier and an aqueous phase. The oil phase is generallycomprised of petrolatum or a fatty alcohol, such as cetyl- or stearylalcohol; the aqueous phase usually, although not necessarily, exceedsthe oil phase in volume, and generally contains a humectant. Theemulsifier in a cream formulation, as explained in Remington: TheScience and Practice of Pharmacy, supra, is generally a nonionic,anionic, cationic or amphoteric surfactant.

Gel formulations can also be used in connection with the presentinvention. As will be appreciated by those working in the field oftopical drug formulation, gels are semisolid. Single-phase gels containorganic macromolecules distributed substantially uniformly throughoutthe carrier liquid, which is typically aqueous, but also may be asolvent or solvent blend.

Ointments, which are semisolid preparations, are typically based onpetrolatum or other petroleum derivatives. As will be appreciated by theordinarily skilled artisan, the specific ointment base to be used is onethat provides for optimum delivery for the active agent chosen for agiven formulation, and, preferably, provides for other desiredcharacteristics as well, e.g., emolliency or the like. As with othercarriers or vehicles, an ointment base should be inert, stable,nonirritating and non-sensitizing. As explained in Remington: TheScience and Practice of Pharmacy, 19th Ed. (Easton, Pa.: Mack PublishingCo., 1995), at pages 1399-1404, ointment bases may be grouped in fourclasses: oleaginous bases; emulsifiable bases; emulsion bases; andwater-soluble bases. Oleaginous ointment bases include, for example,vegetable oils, fats obtained from animals, and semisolid hydrocarbonsobtained from petroleum. Emulsifiable ointment bases, also known asabsorbent ointment bases, contain little or no water and include, forexample, hydroxystearin sulfate, anhydrous lanolin and hydrophilicpetrolatum. Emulsion ointment bases are either water-in-oil (W/O)emulsions or oil-in-water (O/W) emulsions, and include, for example,cetyl alcohol, glyceryl monostearate, lanolin and stearic acid.Preferred water-soluble ointment bases are prepared from polyethyleneglycols of varying molecular weight; again, reference may be had toRemington: The Science and Practice of Pharmacy, supra, for furtherinformation.

Useful formulations of the invention also encompass sprays. Spraysgenerally provide the active agent in an aqueous and/or alcoholicsolution which can be misted onto the skin, nail, hair, claw or hoof fordelivery. Such sprays include those formulated to provide forconcentration of the active agent solution at the site of administrationfollowing delivery, e.g., the spray solution can be primarily composedof alcohol or other like volatile liquid in which the drug or activeagent can be dissolved. Upon delivery to the skin, nail, hair, claw orhoof, the carrier evaporates, leaving concentrated active agent at thesite of administration.

The topical pharmaceutical compositions may also comprise suitable solidor gel phase carriers. Examples of such carriers include but are notlimited to calcium carbonate, calcium phosphate, various sugars,starches, cellulose derivatives, gelatin, and polymers such aspolyethylene glycols.

The topical pharmaceutical compositions may also comprise a suitableemulsifier which refers to an agent that enhances or facilitates mixingand suspending oil-in-water or water-in-oil. The emulsifying agent usedherein may consist of a single emulsifying agent or may be a nonionic,anionic, cationic or amphoteric surfactant or blend of two or more suchsurfactants; preferred for use herein are nonionic or anionicemulsifiers. Such surface-active agents are described in “McCutcheon'sDetergent and Emulsifiers,” North American Edition, 1980 Annualpublished by the McCutcheon Division, MC Publishing Company, 175 RockRoad, Glen Rock, N.J. 07452, USA.

Preferred for use herein are high molecular weight alcohols such ascetearyl alcohol, cetyl alcohol, stearyl alcohol, emulsifying wax,glyceryl monostearate. Other examples are ethylene glycol distearate,sorbitan tristearate, propylene glycol monostearate, sorbitanmonooleate, sorbitan monostearate (SPAN 60), diethylene glycolmonolaurate, sorbitan monopalmitate, sucrose dioleate, sucrose stearate(CRODESTA F-160), polyoxyethylene lauryl ether (BRIJ 30),polyoxyethylene (2) stearyl ether (BRIJ 72), polyoxyethylene (21)stearyl ether (BRIJ 721), polyoxyethylene monostearate (Myrj 45),polyoxyethylene sorbitan monostearate (TWEEN 60), polyoxyethylenesorbitan monooleate (TWEEN 80), polyoxyethylene sorbitan monolaurate(TWEEN 20) and sodium oleate. Cholesterol and cholesterol derivativesmay also be employed in externally used emulsions and promote w/oemulsions.

Especially suitable nonionic emulsifying agents are those withhydrophile-lipophile balances (HLB) of about 3 to 6 for w/o system and 8to 18 for o/w system as determined by the method described by Paul L.Lindner in “Emulsions and Emulsion”, edited by Kenneth Lissant,published by Dekker, New York, N.Y., 1974, pages 188-190. More preferredfor use herein are one or more nonionic surfactants that produce asystem having HLB of about 8 to about 18.

Examples of such nonionic emulsifiers include but are not limited to“BRIJ 72”, the trade name for a polyoxyethylene (2) stearyl ether havingan HLB of 4.9; “BRIJ 721”, the trade name for a polyoxyethylene (21)stearyl ether having an HLB of 15.5, “Brij 30”, the trade name forpolyoxyethylene lauryl ether having an HLB of 9.7; “Polawax”, the tradename for emulsifying wax having an HLB of 8.0; “Span 60”, the trade namefor sorbitan monostearate having an HLB of 4.7; “Crodesta F-160”, thetrade name for sucrose stearate” having an HLB of 14.5. All of thesematerials are available from Ruger Chemicals Inc.; Croda; ICI Americas,Inc.; Spectrum Chemicals; and BASF. When the topical formulations of thepresent invention contain at least one emulsifying agent, eachemulsifying agent is present in amount from about 0.5 to about 2.5 wt %,preferably 0.5 to 2.0%, more preferably 1.0% or 1.8%. Preferably theemulsifying agent comprises a mixture of steareth 21 (at about 1.8%) andsteareth 2 (at about 1.0%).

The topical pharmaceutical compositions may also comprise suitableemollients. Emollients are materials used for the prevention or reliefof dryness, as well as for the protection of the skin, nail, hair, clawor hoof. Useful emollients include, but are not limited to, cetylalcohol, isopropyl myristate, stearyl alcohol, and the like. A widevariety of suitable emollients are known and can be used herein. Seee.g., Sagarin, Cosmetics, Science and Technology, 2nd Edition, Vol. 1,pp. 32-43 (1972), and U.S. Pat. No. 4,919,934, to Deckner et al., issuedApr. 24, 1990, both of which are incorporated herein by reference intheir entirety. These materials are available from Ruger Chemical Co,(Irvington, N.J.).

When the topical formulations of the present invention contain at leastone emollient, each emollient is present in an amount from about 0.1 to15%, preferably 0.1 to about 3.0, more preferably 0.5, 1.0, or 2.5 wt %.Preferably the emollient is a mixture of cetyl alcohol, isopropylmyristate and stearyl alcohol in a 1/5/2 ratio. The emollient may alsobe a mixture of cetyl alcohol and stearyl alcohol in a 1/2 ratio.

The topical pharmaceutical compositions may also comprise suitableantioxidants, substances known to inhibit oxidation. Antioxidantssuitable for use in accordance with the present invention include, butare not limited to, butylated hydroxytoluene, ascorbic acid, sodiumascorbate, calcium ascorbate, ascorbic palmitate, butylatedhydroxyanisole, 2,4,5-trihydroxybutyrophenone,4-hydroxymethyl-2,6-di-tert-butylphenol, erythorbic acid, gum guaiac,propyl gallate, thiodipropionic acid, dilauryl thiodipropionate,tert-butylhydroquinone and tocopherols such as vitamin E, and the like,including pharmaceutically acceptable salts and esters of thesecompounds. Preferably, the antioxidant is butylated hydroxytoluene,butylated hydroxyanisole, propyl gallate, ascorbic acid,pharmaceutically acceptable salts or esters thereof, or mixturesthereof. Most preferably, the antioxidant is butylated hydroxytoluene.These materials are available from Ruger Chemical Co, (Irvington, N.J.).

When the topical formulations of the present invention contain at leastone antioxidant, the total amount of antioxidant present is from about0.001 to 0.5 wt %, preferably 0.05 to about 0.5 wt %, more preferably0.1%.

The topical pharmaceutical compositions may also comprise suitablepreservatives. Preservatives are compounds added to a pharmaceuticalformulation to act as an anti-microbial agent. Among preservatives knownin the art as being effective and acceptable in parenteral formulationsare benzalkonium chloride, benzethonium, chlorohexidine, phenol,m-cresol, benzyl alcohol, methylparaben, propylparaben, chlorobutanol,o-cresol, p-cresol, chlorocresol, phenylmercuric nitrate, thimerosal,benzoic acid, and various mixtures thereof. See, e.g., Wallhausser,K.-H., Develop. Biol. Standard, 24:9-28 (1974) (S. Krager, Basel).Preferably, the preservative is selected from methylparaben,propylparaben and mixtures thereof. These materials are available fromInolex Chemical Co (Philadelphia, Pa.) or Spectrum Chemicals.

When the topical formulations of the present invention contain at leastone preservative, the total amount of preservative present is from about0.01 to about 0.5 wt %, preferably from about 0.1 to 0.5%, morepreferably from about 0.03 to about 0.15. Preferably the preservative isa mixture of methylparaben and proplybarben in a 5/1 ratio. When alcoholis used as a preservative, the amount is usually 15 to 20%.

The topical pharmaceutical compositions may also comprise suitablechelating agents to form complexes with metal cations that do not crossa lipid bilayer. Examples of suitable chelating agents include ethylenediamine tetraacetic acid (EDTA), ethylene glycol-bis(beta-aminoethylether)-N,N,N′,N′-tetraacetic acid (EGTA) and8-Amino-2-[(2-amino-5-methylphenoxy)methyl]-6-methoxyquinoline-N,N,N′,N′-tetraaceticacid, tetrapotassium salt (QUIN-2). Preferably the chelating agents areEDTA and citric acid. These materials are available from SpectrumChemicals.

When the topical formulations of the present invention contain at leastone chelating agent, the total amount of chelating agent present is fromabout 0.005% to 2.0% by weight, preferably from about 0.05% to about 0.5wt %, more preferably about 0.1% by weight.

The topical pharmaceutical compositions may also comprise suitableneutralizing agents used to adjust the pH of the formulation to within apharmaceutically acceptable range. Examples of neutralizing agentsinclude but are not limited to trolamine, tromethamine, sodiumhydroxide, hydrochloric acid, citric acid, and acetic acid. Suchmaterials are available from are available from Spectrum Chemicals(Gardena, Calif.).

When the topical formulations of the present invention contain at leastone neutralizing agent, the total amount of neutralizing agent presentis from about 0.1 wt to about 10 wt %, preferably 0.1 wt % to about 5.0wt %, and more preferably about 1.0 wt %. The neutralizing agent isgenerally added in whatever amount is required to bring the formulationto the desired pH.

The topical pharmaceutical compositions may also comprise suitableviscosity increasing agents. These components are diffusible compoundscapable of increasing the viscosity of a polymer-containing solutionthrough the interaction of the agent with the polymer. CARBOPOL ULTREZ10 may be used as a viscosity-increasing agent. These materials areavailable from Noveon Chemicals, Cleveland, Ohio.

When the topical formulations of the present invention contain at leastone viscosity increasing agent, the total amount of viscosity increasingagent present is from about 0.25% to about 5.0% by weight, preferablyfrom about 0.25% to about 1.0 wt %, and more preferably from about 0.4%to about 0.6% by weight.

The topical pharmaceutical compositions may also comprise suitable nailpenetration enhancers. Examples of nail penetration enhancers includemercaptan compounds, sulfites and bisulfites, keratolytic agents andsurfactants. Nail penetration enhancers suitable for use in theinvention are described in greater detail in Malhotra et al., J. Pharm.Sci., 91:2, 312-323 (2002), which is incorporated herein by reference inits entirety.

The topical pharmaceutical compositions may also comprise one or moresuitable solvents. The ability of any solid substance (solute) todissolve in any liquid substance (solvent) is dependent upon thephysical properties of the solute and the solvent. When solutes andsolvents have similar physical properties the solubility of the solutein the solvent will be the greatest. This gives rise to the traditionalunderstanding that “like dissolves like.” Solvents can be characterizedin one extreme as non-polar, lipophilic oils, while in the other extremeas polar hydrophilic solvents. Oily solvents dissolve other non-polarsubstances by Van der Wals interactions while water and otherhydrophilic solvents dissolve polar substances by ionic, dipole, orhydrogen bonding interactions. All solvents can be listed along acontinuum from the least polar, i.e. hydrocarbons such as decane, to themost polar solvent being water. A solute will have its greatestsolubility in solvents having equivalent polarity. Thus, for drugshaving minimal solubility in water, less polar solvents will provideimproved solubility with the solvent having polarity nearly equivalentto the solute providing maximum solubility. Most drugs have intermediatepolarity, and thus experience maximum solubility in solvents such aspropylene glycol or ethanol, which are significantly less polar thanwater. If the drug has greater solubility in propylene glycol (forexample 8% (w/w)) than in water (for example 0.1% (w/w)), then additionof water to propylene glycol should decrease the maximum amount of drugsolubility for the solvent mixture compared with pure propylene glycol.Addition of a poor solvent to an excellent solvent will decrease themaximum solubility for the blend compared with the maximum solubility inthe excellent solvent.

When compounds are incorporated into topical formulations theconcentration of active ingredient in the formulation may be limited bythe solubility of the active ingredient in the chosen solvent and/orcarrier. Non-lipophilic drugs typically display very low solubility inpharmaceutically acceptable solvents and/or carriers. For example, thesolubility of some compounds in the invention in water is less than0.00025% wt/wt. The solubility of the same compounds in the inventioncan be less than about 2% wt/wt in either propylene glycol or isopropylmyristate. In one embodiment of the present invention, diethylene glycolmonoethyl ether (DGME) is the solvent used to dissolve the compounds ofFormula (I) of Formula (II). The compounds in the invention useful inthe present formulation are believed to have a solubility of from about10% wt/wt to about 25% wt/wt in DGME. In another embodiment a DGME watercosolvent system is used to dissolve the compounds of Formula (I) ofFormula (II). The solvent capacity of DGME drops when water is added;however, the DGME/water cosolvent system can be designed to maintain thedesired concentration of from about 0.1% to about 5% wt/wt activeingredient. Preferably the active ingredient is present from about 0.5%to about 3% wt/wt, and more preferably at about 1% wt/wt, in theas-applied topical formulations. Because DGME is less volatile thanwater, as the topical formulation evaporates upon application, theactive agent becomes more soluble in the cream formulation. Thisincreased solubility reduces the likelihood of reduced bioavailabilitycaused by the drug precipitating on the surface of the skin, nail, hair,claw or hoof.

Liquid forms, such as lotions suitable for topical administration orsuitable for cosmetic application, may include a suitable aqueous ornonaqueous vehicle with buffers, suspending and dispensing agents,thickeners, penetration enhancers, and the like. Solid forms such ascreams or pastes or the like may include, for example, any of thefollowing ingredients, water, oil, alcohol or grease as a substrate withsurfactant, polymers such as polyethylene glycol, thickeners, solids andthe like. Liquid or solid formulations may include enhanced deliverytechnologies such as liposomes, microsomes, microsponges and the like.

Additionally, the compounds can be delivered using a sustained-releasesystem, such as semipermeable matrices of solid hydrophobic polymerscontaining the therapeutic agent. Various sustained-release materialshave been established and are well known by those skilled in the art.

Topical treatment regimens according to the practice of this inventioncomprise applying the composition directly to the skin, nail, hair, clawor hoof at the application site, from one to several times daily.

Formulations of the present invention can be used to treat, ameliorateor prevent conditions or symptoms associated with bacterial infections,acne, inflammation and the like.

In an exemplary embodiment, the pharmaceutical formulation includes asimple solution. In an exemplary embodiment, the simple solutionincludes an alcohol. In an exemplary embodiment, the simple solutionincludes alcohol and water. In an exemplary embodiment, the alcohol isethanol, ethylene glycol, propanol, polypropylene glycol, isopropanol orbutanol. In another exemplary embodiment, the simple solution is amember selected from about 10% polypropylene glycol and about 90%ethanol; about 20% polypropylene glycol and about 80% ethanol; about 30%polypropylene glycol and about 70% ethanol; about 40% polypropyleneglycol and about 60% ethanol; about 50% polypropylene glycol and about50% ethanol; about 60% polypropylene glycol and about 40% ethanol; about70% polypropylene glycol and about 30% ethanol; about 80% polypropyleneglycol and about 20% ethanol; about 90% polypropylene glycol and about10% ethanol.

In an exemplary embodiment, the pharmaceutical formulation is a lacquer.Please see Remington's, supra, for more information on the production oflacquers.

In an exemplary embodiment, the compound is present in saidpharmaceutical formulation in a concentration of from about 0.5% toabout 15%. In an exemplary embodiment, the compound is present in saidpharmaceutical formulation in a concentration of from about 0.1% toabout 12.5%. In an exemplary embodiment, the compound is present in saidpharmaceutical formulation in a concentration of from about 1% to about10%. In an exemplary embodiment, the compound is present in saidpharmaceutical formulation in a concentration of from about 1% to about5%. In an exemplary embodiment, the compound is present in saidpharmaceutical formulation in a concentration of from about 2% to about8%. In an exemplary embodiment, the compound is present in saidpharmaceutical formulation in a concentration of from about 4% to about9%.

VII. b) Additional Active Agents

The following are examples of the cosmetic and pharmaceutical agentsthat can be added to the topical pharmaceutical formulations of thepresent invention. The following agents are known compounds and arereadily available commercially.

Anti-inflammatory agents include, but are not limited to, bisabolol,mentholatum, dapsone, aloe, hydrocortisone, and the like.

Vitamins include, but are not limited to, Vitamin B, Vitamin E, VitaminA, Vitamin D, and the like and vitamin derivatives such as tazarotene,calcipotriene, tretinoin, adapalene and the like.

Anti-aging agents include, but are not limited to, niacinamide, retinoland retinoid derivatives, AHA, Ascorbic acid, lipoic acid, coenzyme Q10, beta hydroxy acids, salicylic acid, copper binding peptides,dimethylaminoethyl (DAEA), and the like.

Sunscreens and or sunburn relief agents include, but are not limited to,PABA, jojoba, aloe, padimate-O, methoxycinnamates, proxamine HCl,lidocaine and the like. Sunless tanning agents include, but are notlimited to, dihydroxyacetone (DHA).

Psoriasis-treating agents and/or acne-treating agents include, but arenot limited to, salicylic acid, benzoyl peroxide, coal tar, seleniumsulfide, zinc oxide, pyrithione (zinc and/or sodium), tazarotene,calcipotriene, tretinoin, adapalene and the like.

Agents that are effective to control or modify keratinization, includingwithout limitation: tretinoin, tazarotene, and adapalene.

The compositions comprising an compound/active agent of Formula (I) ofFormula (II), and optionally at least one of these additional agents,are to be administered topically. In a primary application, this leadsto the compounds of the invention and any other active agent workingupon and treating the skin, nail, hair, claw or hoof. Alternatively, anyone of the topically applied active agents may also be deliveredsystemically by transdermal routes.

In such compositions an additional cosmetically or pharmaceuticallyeffective agent, such as an anti-inflammatory agent, vitamin, anti-agingagent, sunscreen, and/or acne-treating agent, for example, is usually aminor component (from about 0.001% to about 20% by weight or preferablyfrom about 0.01% to about 10% by weight) with the remainder beingvarious vehicles or carriers and processing aids helpful for forming thedesired dosing form.

VII. c) Testing

Preferred compounds for use in the present topical formulations willhave certain pharmacological properties. Such properties include, butare not limited to, low toxicity, low serum protein binding anddesirable in vitro and in vivo half-lives. Assays may be used to predictthese desirable pharmacological properties. Assays used to predictbioavailability include transport across human intestinal cellmonolayers, including Caco-2 cell monolayers. Serum protein binding maybe predicted from albumin binding assays. Such assays are described in areview by Oravcova et al. (1996, J. Chromat. B677: 1-27). Compoundhalf-life is inversely proportional to the frequency of dosage of acompound. In vitro half-lives of compounds may be predicted from assaysof microsomal half-life as described by Kuhnz and Gleschen (DrugMetabolism and Disposition, (1998) volume 26, pages 1120-1127).

Toxicity and therapeutic efficacy of such compounds can be determined bystandard pharmaceutical procedures in cell cultures or experimentalanimals, e.g., for determining the LD50 (the dose lethal to 50% of thepopulation) and the ED₅₀ (the dose therapeutically effective in 50% ofthe population). The dose ratio between toxic and therapeutic effects isthe therapeutic index and it can be expressed as the ratio between LD₅₀and ED₅₀. Compounds that exhibit high therapeutic indices are preferred.The data obtained from these cell culture assays and animal studies canbe used in formulating a range of dosage for use in humans. The dosageof such compounds lies preferably within a range of circulatingconcentrations that include the ED₅₀ with little or no toxicity. Thedosage can vary within this range depending upon the dosage formemployed and the route of administration utilized. The exactformulation, route of administration and dosage can be chosen by theindividual physician in view of the patient's condition. (See, e.g.Fingl et al., 1975, in “The Pharmacological Basis of Therapeutics”, Ch.1, p. 1).

VII. d) Administration

For any compound used in the method of the invention, thetherapeutically effective dose can be estimated initially from cellculture assays, as disclosed herein. For example, a dose can beformulated in animal models to achieve a circulating concentration rangethat includes the EC₅₀ (effective dose for 50% increase) as determinedin cell culture, i.e., the concentration of the test compound whichachieves a half-maximal inhibition of bacterial cell growth. Suchinformation can be used to more accurately determine useful doses inhumans.

In general, the compounds prepared by the methods, and from theintermediates, described herein will be administered in atherapeutically or cosmetically effective amount by any of the acceptedmodes of administration for agents that serve similar utilities. It willbe understood, however, that the specific dose level for any particularpatient will depend upon a variety of factors including the activity ofthe specific compound employed, the age, body weight, general health,sex, diet, time of administration, route of administration, and rate ofexcretion, drug combination, the severity of the particular diseaseundergoing therapy and the judgment of the prescribing physician. Thedrug can be administered from once or twice a day, or up to 3 or 4 timesa day.

Dosage amount and interval can be adjusted individually to provideplasma levels of the active moiety that are sufficient to maintainbacterial cell growth inhibitory effects. Usual patient dosages forsystemic administration range from 0.1 to 1000 mg/day, preferably, 1-500mg/day, more preferably 10-200 mg/day, even more preferably 100-200mg/day. Stated in terms of patient body surface areas, usual dosagesrange from 50-91 mg/m²/day.

The amount of the compound in a formulation can vary within the fullrange employed by those skilled in the art. Typically, the formulationwill contain, on a weight percent (wt %) basis, from about 0.01-10 wt %of the drug based on the total formulation, with the balance being oneor more suitable pharmaceutical excipients. Preferably, the compound ispresent at a level of about 0.1-3.0 wt %, more preferably, about 1.0 wt%.

The invention is further illustrated by the Examples that follow. TheExamples are not intended to define or limit the scope of the invention.

EXAMPLES

Proton NMR are recorded on Varian AS 300 spectrometer and chemicalshifts are reported as δ (ppm) down field from tetramethylsilane. Massspectra are determined on Micromass Quattro II.

Example 1 Preparation of 3 from 1

1.1 Reduction of Carboxylic Acid

To a solution of 1 (23.3 mmol) in anhydrous THF (70 mL) under nitrogenwas added dropwise a BH₃ THF solution (1.0 M, 55 mL, 55 mmol) at 0° C.and the reaction mixture was stirred overnight at room temperature. Thenthe mixture was cooled again with ice bath and MeOH (20 mL) was addeddropwise to decompose excess BH₃. The resulting mixture was stirreduntil no bubble was released and then 10% NaOH (10 mL) was added. Themixture was concentrated and the residue was mixed with water (200 mL)and extracted with EtOAc. The residue from rotary evaporation waspurified by flash column chromatography over silica gel to give 20.7mmol of 3.

1.2 Results

Exemplary compounds of structure 3 prepared by the method above areprovided below.

1.2.a 2-Bromo-5-chlorobenzyl Alcohol

¹H NMR (300 MHz, DMSO-d₆): δ 7.57 (d, J=8.7 Hz, 1H), 7.50-7.49 (m, 1H),7.28-7.24 (m, 1H), 5.59 (t, J=6.0 Hz, 1H) and 4.46 (d, J=6.0 Hz, 2H)ppm.

1.2.b 2-Bromo-5-methoxybenzyl Alcohol

¹H NMR (300 MHz, DMSO-d₆): δ 7.42 (d, J=8.7 Hz, 1H), 7.09 (d, J=2.4 Hz,1H), 6.77 (dd, J₁=3 Hz, J₂=3 Hz, 1H), 5.43 (t, J=5.7 Hz, 1H), 4.44 (d,J=5.1 Hz, 2H), 3.76 (s, 3H).

Example 2 Preparation of 3 from 2

2.1. Reduction of Aldehyde

To a solution of 2 (Z═H, 10.7 mmol) in methanol (30 mL) was added sodiumborohydride (5.40 mol), and the mixture was stirred at room temperaturefor 1 h. Water was added, and the mixture was extracted with ethylacetate. The organic layer was washed with brine and dried on anhydroussodium sulfate. The solvent was removed under reduced pressure to afford9.9 mmol of 3.

2.2 Results

Exemplary compounds of structure 3 prepared by the method above areprovided below.

2.2.a 2-Bromo-5-(4-cyanophenoxy)benzyl Alcohol

¹H-NMR (300 MHz, CDCl₃) δ (ppm) 2.00 (br s, 1H), 4.75 (s, 2H), 6.88 (dd,J=8.5, 2.9 Hz, 1H), 7.02 (d, J=8.8 Hz, 1H), 7.26 (d, J=2.6 Hz, 1H), 7.56(d, J=8.5 Hz, 1H), 7.62 (d, J=8.8 Hz, 2H).

2.2.b 2-Bromo-4-(4-cyanophenoxy)benzyl Alcohol

¹H NMR (300 MHz, DMSO-d₆): δ 7.83 (d, 2H), 7.58 (d, 1H), 7.39 (d, 1H),7.18 (dd, 1H), 7.11 (d, 2H), 5.48 (t, 1H) and 4.50 (d, 2H) ppm.

2.2.c 5-(4-Cyanophenoxy)-1-Indanol

M.p. 50-53° C. MS (ESI+): m/z=252 (M+1). HPLC: 99.7% purity at 254 nmand 99.0% at 220 nm. ¹H NMR (300 MHz, DMSO-d₆): δ 7.80 (d, 2H), 7.37 (d,1H), 7.04 (d, 2H), 6.98-6.93 (m, 2H), 5.27 (d, 1H), 5.03 (q, 1H),2.95-2.85 (m, 1H), 2.75-2.64 (m, 1H), 2.39-2.29 (m, 1H) and 1.85-1.74(m, 1H) ppm.

2.2.d 2-Bromo-5-(tert-butyldimethylsiloxy)benzyl Alcohol

¹H-NMR (300 MHz, CDCl₃) δ (ppm) 0.20 (s, 6H), 0.98 (s, 9H), 4.67 (br s,1H), 6.65 (dd, J=8.2, 2.6 Hz, 1H), 6.98 (d, J=2.9 Hz, 1H), 7.36 (d,J=8.8 Hz, 1H).

Additional examples of compounds which can be produced by this methodinclude 2-bromo-4-(3-cyanophenoxy)benzyl alcohol;2-bromo-4-(4-chlorophenoxy)benzyl alcohol; 2-bromo-4-phenoxybenzylalcohol; 2-bromo-5-(3,4-dicyanophenoxy)benzyl alcohol;2-(2-bromo-5-fluorophenyl)ethyl alcohol; 2-bromo-5-fluorobenzyl alcohol;and 1-bromo-2-naphthalenemethanol.

Example 3 Preparation of 4 from 3

3.1 Protective Alkylation

Compound 3 (20.7 mmol) was dissolved in CH₂Cl₂ (150 mL) and cooled to 0°C. with ice bath. To this solution under nitrogen were added in sequenceN,N-di-isopropyl ethyl amine (5.4 mL, 31.02 mmol, 1.5 eq) andchloromethyl methyl ether (2 mL, 25.85 mmol, 1.25 eq). The reactionmixture was stirred overnight at room temperature and washed withNaHCO₃-saturated water and then NaCl-saturated water. The residue afterrotary evaporation was purified by flash column chromatography oversilica gel to give 17.6 mmol of 4.

3.2 Results

Exemplary compounds of structure 4 prepared by the method above areprovided below.

3.2.a 2-Bromo-5-chloro-1-(methoxymethoxymethyl)benzene

¹H NMR (300 MHz, DMSO-d₆): δ 7.63 (d, J=8.7 Hz, 1H), 7.50 (dd, J=2.4 &0.6 Hz, 1H), 7.32 (dd, J=8.4 & 2.4 Hz, 1H), 4.71 (s, 2H), 4.53 (s, 2H)and 3.30 (s, 3H) ppm.

3.2.b 2-Bromo-5-fluoro-1-[1-(methoxymethoxy)ethyl]benzene

¹H-NMR (300.058 MHz, CDCl₃) δ ppm 1.43 (d, J=6.5 Hz, 3H), 3.38 (s, 3H),4.55 (d, J=6.5 Hz, 1H), 4.63 (d, J=6.5 Hz, 1H), 5.07 (q, J=6.5 Hz, 1H),6.85 (m, 1H), 7.25 (dd, J=9.7, 2.6 Hz, 1H), 7.46 (dd, J=8.8, 5.3 Hz,1H).

3.2.c 2-Bromo-5-fluoro-1-[2-(methoxymethoxy)ethyl]benzene

¹H-NMR (300.058 MHz, CDCl₃) δ ppm 3.04 (t, J=6.7 Hz, 2H), 3.31 (s, 3H),3.77 (t, J=6.7 Hz, 2H), 4.62 (s, 2H), 6.82 (td, J=8.2, 3.2 Hz, 1H), 7.04(dd, J=9.4, 2.9 Hz, 1H), 7.48 (dd, J=8.8, 5.3 Hz, 1H).

3.2.d 2-Bromo-4,5-difluoro-1-(methoxymethoxymethyl)benzene

¹H-NMR (300.058 MHz, CDCl₃) δ ppm 3.42 (s, 3H), 4.57 (d, J=1.2 Hz, 2H),4.76 (s, 2H), 7.3-7.5 (m, 2H).

3.2.e 2-Bromo-5-cyano-1-(methoxymethoxymethyl)benzene

¹H-NMR (300.058 MHz, CDCl₃) δ ppm 3.43 (s, 3H), 4.65 (s, 2H), 4.80 (s,2H), 7.43 (dd, J=8.2, 4.1 Hz, 1H), 7.66 (d, J=8.2 Hz, 1H), 7.82 (d,J=4.1 Hz, 1H).

3.2.f 2-Bromo-5-methoxy-1-(methoxymethoxymethyl)benzene

¹H NMR (300 MHz, DMSO-d₆): δ 7.48 (dd, J₁=1.2 Hz, J₂=1.2 Hz, 1H), 7.05(d, J=2.7 Hz, 1H), 6.83 (dd, J₁=3 Hz, J₂=3 Hz, 1H), 4.69 (d, J=1.2 Hz,2H), 4.5 (s, 2H), 3.74 (d, J=1.5 Hz, 3H), 3.32 (d, J=2.1 Hz, 3H) ppm.

3.2.g 1-Benzyl-1-(2-bromophenyl)-1-(methoxymethoxy)ethane

¹H NMR (300 MHz, DMSO-d₆): δ 7.70-7.67 (m, 1H), 7.25-7.09 (m, 6H),6.96-6.93 (m, 2H), 4.61 (d, 1H), 4.48 (d, 1H), 3.36-3.26 (m, 2H), 3.22(s, 3H) and 1.63 (s, 3H) ppm.

3.2.h 2-Bromo-6-fluoro-1-(methoxymethoxymethyl)benzene

¹H-NMR (300 MHz, CDCl₃) δ (ppm) 3.43 (s, 3H), 4.74 (s, 2H), 4.76 (d,J=2.1 Hz, 2H), 7.05 (t, J=9.1 Hz, 1H), 7.18 (td, J=8.2, 5.9 Hz, 1H),7.40 (d, J=8.2 Hz, 1H).

3.2.i 2-Bromo-4-(4-cyanophenoxy)-1-(methoxymethoxymethyl)benzene

¹H NMR (300 MHz, DMSO-d₆): δ 7.84 (d, 2H), 7.56 (d, 1H), 7.44 (d, 1H),7.19-7.12 (m, 3H), 4.69 (s, 2H), 4.56 (s, 2H) and 3.31 (s, 3H) ppm.

3.2.j2-Bromo-5-(tert-butyldimethylsiloxy)-1-(methoxymethoxymethyl)benzene

¹H-NMR (300 MHz, CDCl₃) δ (ppm) 0.19 (s, 6H), 0.98 (s, 9H), 3.43 (s,3H), 4.59 (s, 2H), 4.75 (s, 2H), 6.64 (dd, J=8.5, 2.9 Hz, 1H), 6.98 (d,J=2.9 Hz, 1H), 7.36 (d, J=8.5 Hz, 1H).

3.2.k 2-Bromo-5-(2-cyanophenoxy)-1-(methoxymethoxymethyl)benzene

¹H-NMR (300 MHz, CDCl₃) δ (ppm) 3.41 (s, 3H), 4.64 (s, 2H), 4.76 (s,2H), 6.8-6.9 (m, 2H), 7.16 (td, J=7.6, 0.9 Hz, 1H), 7.28 (d, J=2.9 Hz,1H), 7.49 (ddd, J=8.8, 7.6, 1.8 Hz, 1H), 7.56 (d, J=8.5 Hz, 1H), 7.67(dd, J=7.9, 1.8 Hz, 1H).

3.2.l 2-Bromo-5-phenoxy-1-(methoxymethoxymethyl)benzene

¹H-NMR (300 MHz, CDCl₃) δ (ppm) 3.40 (s, 3H), 4.62 (s, 2H), 4.74 (s,2H), 6.80 (dd, J=8.8, 2.9 hz, 1H), 7.01 (d, J=8.5 Hz, 2H), 7.12 (t,J=7.9 Hz, 1H), 7.19 (d, J=2.9 hz, 1H), 7.35 (t, J=7.6 Hz, 2H), 7.48 (d,J=8.5 Hz, 1H).

Additional examples of compounds which can be produced by this methodinclude 2-bromo-1-(methoxymethoxymethyl)benzene;2-bromo-5-methyl-1-(methoxymethoxymethyl)benzene;2-bromo-5-(methoxymethoxymethyl)-1-(methoxymethoxymethyl)benzene;2-bromo-5-fluoro-1-(methoxymethoxymethyl)benzene;1-bromo-2-(methoxymethoxymethyl)naphthalene;2-bromo-4-fluoro-1-(methoxymethoxymethyl)benzene;2-phenyl-1-(2-bromophenyl)-1-(methoxymethoxy)ethane;2-bromo-5-(4-cyanophenoxy)-1-(methoxymethoxy methyl)benzene;2-bromo-4-(3-cyanophenoxy)-1-(methoxymethoxymethyl)benzene;2-bromo-4-(4-chlorophenoxy)-1-(methoxymethoxymethyl)benzene;2-bromo-4-phenoxy-1-(methoxymethoxymethyl)benzene;2-bromo-5-(3,4-dicyanophenoxy)-1-(methoxymethoxymethyl)benzene.

Example 4 Preparation of I from 4 Via 5

4.1 Metallation and Boronylation

To a solution of 4 (17.3 mmol) in anhydrous THF (80 mL) at −78° C. undernitrogen was added dropwise tert-BuLi or n-BuLi (11.7 mL) and thesolution became brown colored. Then, B(OMe)₃ (1.93 mL, 17.3 mmol) wasinjected in one portion and the cooling bath was removed. The mixturewas warmed gradually with stirring for 30 min and then stirred with awater bath for 2 h. After addition of 6N HCl (6 mL), the mixture wasstirred overnight at room temperature and about 50% hydrolysis hashappened as shown by TLC analysis. The solution was rotary evaporatedand the residue was dissolved in MeOH (50 mL) and 6N HCl (4 mL). Thesolution was refluxed for 1 h and the hydrolysis was completed asindicated by TLC analysis. Rotary evaporation gave a residue which wasdissolved in EtOAc, washed with water, dried and then evaporated. Thecrude product was purified by flash column chromatography over silicagel to provide a solid with 80% purity. The solid was further purifiedby washing with hexane to afford 7.2 mmol of I.

4.2 Results

Analytical data for exemplary compounds of structure I are providedbelow.

4.2.a 5-Chloro-1,3-dihydro-1-hydroxy-2,1-benzoxaborole (C1)

M.p. 142-150° C. MS (ESI): m/z=169 (M+1, positive) and 167 (M−1,negative). HPLC (220 nm): 99% purity. ¹H NMR (300 MHz, DMSO-d₆): δ 9.30(s, 1H), 7.71 (d, J=7.8 Hz, 1H), 7.49 (s, 1H), 7.38 (d, J=7.8 Hz, 1H)and 4.96 (s, 2H) ppm.

4.2.b 1,3-Dihydro-1-hydroxy-2,1-benzoxaborole (C2)

M.p. 83-86° C. MS (ESI): m/z=135 (M+1, positive) and 133 (M−1,negative). HPLC (220 nm): 95.4% purity. ¹H NMR (300 MHz, DMSO-d₆): δ9.14 (s, 1H), 7.71 (d, J=7.2 Hz, 1H), 7.45 (t, J=7.5 Hz, 1H), 7.38 (d,J=7.5 Hz, 1H), 7.32 (t, J=7.1 Hz, 1H) and 4.97 (s, 2H) ppm.

4.2.c 5-Fluoro-1,3-dihydro-1-hydroxy-3-methyl-2,1-benzoxaborole (C3)

¹H-NMR (300 MHz, DMSO-d₆) δ ppm 1.37 (d, J=6.4 Hz, 3H), 5.17 (q, J=6.4Hz, 1H), 7.14 (m, 1H), 7.25 (dd, J=9.7, 2.3 Hz, 1H), 7.70 (dd, J=8.2,5.9 Hz, 1H), 9.14 (s, 1H).

4.2.d 6-Fluoro-1-hydroxy-1,2,3,4-tetrahydro-2,1-benzoxaborine (C4)

¹H-NMR (300 MHz, DMSO-d₆) δ ppm 2.86 (t, J=5.9 Hz, 2H), 4.04 (t, J=5.9Hz, 2H), 7.0-7.1 (m, 2H), 7.69 (dd, J=8.2, 7.2 Hz, 1H), 8.47 (s, 1H).

4.2.e 5,6-Difluoro-1,3-dihydro-1-hydroxy-2,1-benzoxaborole (C5)

¹H-NMR (300 MHz, DMSO-d₆) δ ppm 4.94 (s, 2H), 7.50 (dd, J=10.7, 6.8 Hz,1H), 7.62 (dd, J=9.7, 8.2 Hz, 1H), 9.34 (s, 1H).

4.2.f 5-Cyano-1,3-dihydro-1-hydroxy-2,1-benzoxaborole (C6)

¹H-NMR (300 MHz, DMSO-d₆) δ ppm 5.03 (s, 2H), 7.76 (d, J=8.2 Hz, 1H),7.89 (d, J=8.2 Hz, 1H), 7.90 (s, 1H), 9.53 (s, 1H).

4.2.g 1,3-Dihydro-1-hydroxy-5-methoxy-2,1-benzoxaborole (C7)

M.p. 102-104° C. MS ESI: m/z=165.3 (M+1) and 162.9 (M−1). ¹H NMR (300MHz, DMSO-d₆): δ 8.95 (s, 1H), 7.60 (d, J=8.1 Hz, 1H), 6.94 (s, 1H),6.88 (d, J=8.1 Hz, 1H), 4.91 (s, 2H), 3.77 (s, 3 H) ppm.

4.2.h 1,3-Dihydro-1-hydroxy-5-methyl-2,1-benzoxaborole (C8)

M.p. 124-128° C. MS ESI: m/z=148.9 (M+1) and 146.9 (M−1). ¹H NMR (300MHz, DMSO-d₆): δ 9.05 (s, 1H), 7.58 (d, J=7.2 Hz, 1H), 7.18 (s, 1H),7.13 (d, J=7.2 Hz, 2H), 4.91 (s, 2H), 2.33 (s, 3H) ppm.

4.2.i 1,3-Dihydro-1-hydroxy-5-hydroxymethyl-2,1-benzoxaborole (C9)

MS: m/z=163 (M−1, ESI−). ¹H NMR (300 MHz, DMSO-d₆): δ 9.08 (s, 1H), 7.64(d, 1H), 7.33 (s, 1H), 7.27 (d, 1H), 5.23 (t, 1H), 4.96 (s, 2H), 4.53(d, 2H) ppm.

4.2.j 1,3-Dihydro-5-fluoro-1-hydroxy-2,1-benzoxaborole (C10)

M.p. 110-114° C. MS ESI: m/z=150.9 (M−1). ¹H NMR (300 MHz, DMSO-d₆): δ9.20 (s, 1H), 7.73 (dd, J₁=6 Hz, J₂=6 Hz, 1H), 7.21 (m, 1H), 7.14 (m,1H), 4.95 (s, 2H) ppm.

4.2.k 1,3-Dihydro-2-oxa-1-cyclopenta[{acute over (α)}]naphthalene (C11)

M.P. 139-143° C. MS ESI: m/z=184.9 (M+1). ¹H NMR (300 MHz, DMSO-d₆): δ9.21 (s, 1H), 8.28 (dd, J₁=6.9 Hz, J₂=0.6 Hz, 1H), 7.99 (d, J=8.1 Hz,1H), 7.95 (d, J=7.5 Hz, 1H), 7.59-7.47 (m, 3H), 5.09 (s, 2H) ppm.

4.2.l 7-Hydroxy-2,1-oxaborolano[5,4-c]pyridine (C12)

¹H-NMR (300 MHz, DMSO-d₆): δ ppm 5.00 (s, 2H), 7.45 (d, J=5.0 Hz, 1H),8.57 (d, J=5.3 Hz, 1H), 8.91 (s, 1H), 9.57 (s, 1H). ESI-MS m/z 134(M-H)⁻, C₆H₆BNO₂=135.

4.2.m 1,3-Dihydro-6-fluoro-1-hydroxy-2,1-benzoxaborole (C13)

M.p. 110-117.5° C. MS (ESI): m/z=151 (M−1, negative). HPLC (220 nm):100% purity. ¹H NMR (300 MHz, DMSO-d₆): δ 9.29 (s, 1H), 7.46-7.41 (m,2H), 7.29 (td, 1H) and 4.95 (s, 2H) ppm.

4.2.n 3-Benzyl-1,3-dihydro-1-hydroxy-3-methyl-2,1-benzoxaborole (C14)

MS (ESI): m/z=239 (M+1, positive). HPLC: 99.5% purity at 220 nm and95.9% at 254 nm. ¹H NMR (300 MHz, DMSO-d₆): δ 8.89 (s, 1H), 7.49-7.40(m, 3H), 7.25-7.19 (m, 1H), 7.09-7.05 (m, 3H), 6.96-6.94 (m, 2H), 3.10(d, 1H), 3.00 (d, 1H) and 1.44 (s, 3H) ppm.

4.2.o 3-Benzyl-1,3-dihydro-1-hydroxy-2,1-benzoxaborole (C15)

MS (ESI+): m/z=225 (M+1). HPLC: 93.4% purity at 220 nm. ¹H NMR (300 MHz,DMSO-d₆): δ 9.08 (s, 1H), 7.63 (dd, 1H), 7.43 (t, 1H), 7.35-7.14 (m,7H), 5.38 (dd, 1H), 3.21 (dd, 1H) and 2.77 (dd, 1H) ppm.

4.2.p 1,3-Dihydro-4-fluoro-1-hydroxy-2,1-benzoxaborole (C16)

¹H-NMR (300 MHz, DMSO-d₆) δ (ppm) 5.06 (s, 2H), 7.26 (ddd, J=9.7, 7.9,0.6 Hz, 1H), 7.40 (td, J=8.2, 4.7 Hz, 1H), 7.55 (d, J=7.0 Hz, 1H), 9.41(s, 1H).

4.2.q 5-(4-Cyanophenoxy)-1,3-dihydro-1-hydroxy-2,1-benzoxaborole (C17)

¹H-NMR (300 MHz, DMSO-d₆) δ ppm 4.95 (s, 2H), 7.08 (dd, J=7.9, 2.1 Hz,1H), 7.14 (d, J=8.8 Hz, 1H), 7.15 (d, J=2.1 Hz, 1H), 7.78 (d, J=7.9 Hz,1H), 7.85 (d, J=9.1 Hz, 2H), 9.22 (s, 1H).

4.2.r 6-(4-Cyanophenoxy)-1,3-dihydro-1-hydroxy-2,1-benzoxaborole (C18)

M.p. 148-151° C. MS: m/z=252 (M+1) (ESI+) and m/z=250 (M−1) (ESI−).HPLC: 100% purity at 254 nm and 98.7% at 220 nm. ¹H NMR (300 MHz,DMSO-d₆): δ 9.26 (s, 1H), 7.82 (d, 2H), 7.50 (d, 1H), 7.39 (d, 1H), 7.26(dd, 1H), 7.08 (d, 2H) and 4.99 (s, 2H) ppm

4.2.s 6-(3-Cyanophenoxy)-1,3-dihydro-1-hydroxy-2,1-benzoxaborole (C19)

M.p. 146-149° C. MS: m/z=252 (M+1) (ESI+) and m/z=250 (M−1) (ESI−).HPLC: 100% purity at 254 nm and 97.9% at 220 nm. ¹H NMR (300 MHz,DMSO-d₆): δ 9.21 (s, 1H), 7.60-7.54 (m, 2H), 7.50-7.45 (m, 2H),7.34-7.30 (m, 2H), 7.23 (dd, 1H) and 4.98 (s, 2H) ppm.

4.2.t 6-(4-Chlorophenoxy)-1,3-dihydro-1-hydroxy-2,1-benzoxaborole (C20)

M.p. 119-130° C. MS: m/z=261 (M+1) (ESI+) and m/z=259 (M−1) (ESI−).HPLC: 100% purity at 254 nm and 98.9% at 220 nm. ¹H NMR (300 MHz,DMSO-d₆): δ 9.18 (s, 1H), 7.45-7.41 (m, 3H), 7.29 (d, 1H), 7.19 (dd,1H), 7.01 (d, 2H) and 4.96 (s, 2H) ppm.

4.2.u 6-Phenoxy-1,3-dihydro-1-hydroxy-2,1-benzoxaborole (C21)

M.p. 95-99° C. MS: m/z=227 (M+1) (ESI+) and m/z=225 (M−1) (ESI−). HPLC:100% purity at 254 nm and 98.4% at 220 nm. ¹H NMR (300 MHz, DMSO-d₆): δ9.17 (s, 1H), 7.43-7.35 (m, 3H), 7.28 (s, 1H), 7.19-7.09 (m, 2H), 6.99(d, 2H) and 4.96 (s, 2H) ppm.

4.2.v 5-(4-Cyanobenzyloxy)-1,3-dihydro-1-hydroxy-2,1-benzoxaborole (C22)

¹H-NMR (300 MHz, DMSO-d₆) δ (ppm) 4.90 (s, 2H), 5.25 (s, 2H), 6.98 (dd,J=7.9, 2.1 Hz, 1H), 7.03 (d, J=1.8 Hz, 1H), 7.62 (d, J=7.9 Hz, 1H), 7.64(d, J=8.5 Hz, 2H), 7.86 (d, J=8.5 Hz, 1H), 9.01 (s, 1H).

4.2.w 5-(2-Cyanophenoxy)-1,3-dihydro-1-hydroxy-2,1-benzoxaborole (C23)

¹H-NMR (300 MHz, DMSO-d₆) δ (ppm) 4.95 (s, 2H), 7.0-7.2 (m, 3H), 7.32(td, J=7.6, 1.2 Hz, 1H), 7.68 (ddd, J=9.1, 7.6, 1.8 Hz, 1H), 7.77 (d,J=7.9 Hz, 1H), 7.91 (dd, J=7.9, 1.8 Hz, 1H).

4.2.x 5-Phenoxy-1,3-dihydro-1-hydroxy-2,1-benzoxaborole (C24)

¹H-NMR (300 MHz, DMSO-d₆) δ (ppm) 4.91 (s, 2H), 6.94 (s, 1H), 6.96 (d,J=8.8 Hz, 1H), 7.05 (d, J=7.6 Hz, 2H), 7.17 (t, J=7.3 Hz, 1H), 7.41 (t,J=7.3 Hz, 2H), 7.70 (d, J=8.5 Hz, 1H), 9.11 (s, 1H).

4.2.y5-[4-(N,N-Diethylcarbamoyl)phenoxy]-1,3-dihydro-1-hydroxy-2,1-benzoxaborole(C25)

¹H-NMR (300 MHz, DMSO-d₆) δ (ppm) 1.08 (br s, 6H), 3.1-3.5 (m, 4H), 4.93(s, 2H), 7.0-7.1 (m, 4H), 7.37 (d, J=8.5 Hz, 2H), 7.73 (d, J=7.9 Hz,1H), 9.15 (s, 1H).

4.2.z1,3-Dihydro-1-hydroxy-5-[4-(morpholinocarbonyl)phenoxy]-2,1-benzoxaborole(C26)

¹H-NMR (300 MHz, DMSO-d₆) δ (ppm) 3.3-3.7 (m, 8H), 4.93 (s, 2H), 7.0-7.1(m, 4H), 7.44 (d, J=8.8 Hz, 2H), 7.73 (d, J=7.9 Hz, 1H), 9.16 (s, 1H).

4.2.aa 5-(3,4-Dicyanophenoxy)-1,3-dihydro-1-hydroxy-2,1-benzoxaborole(C27)

¹H-NMR (300 MHz, DMSO-d₆) δ (ppm) 4.97 (s, 2H), 7.13 (dd, J=7.9, 2.1 Hz,1H), 7.21 (d, J=1.5 Hz, 1H), 7.43 (dd, J=8.8, 2.6 Hz, 1H), 7.81 (d,J=7.9 Hz, 1H), 7.82 (d, J=2.6 Hz, 1H), 8.11 (d, J=8.5 Hz, 1H), 9.26 (s,1H).

4.2.ab 6-Phenylthio-1,3-dihydro-1-hydroxy-2,1-benzoxaborole (C28)

M.p. 121-124° C. MS: m/z=243 (M+1) (ESI+) and m/z=241 (M−1) (ESI−).HPLC: 99.6% purity at 254 nm and 99.6% at 220 nm. ¹H NMR (300 MHz,DMSO-d₆): δ 9.25 (s, 1H), 7.72 (dd, 1H), 7.48 (dd, 1H), 7.43 (dd, 1H),7.37-7.31 (m, 2H), 7.29-7.23 (m, 3H), and 4.98 (s, 2H) ppm.

4.2.ac6-(4-trifluoromethoxyphenoxy)-1,3-dihydro-1-hydroxy-2,1-benzoxaborole(C29)

M.p. 97-101° C. MS: m/z=311 (M+1) (ESI+) and m/z=309 (M−1) (ESI−). HPLC:100% purity at 254 nm and 100% at 220 nm. ¹H NMR (300 MHz, DMSO-d₆): δ9.20 (s, 1H), 7.45 (d, 1H), 7.37 (d, 2H), 7.33 (d, 1H), 7.21 (dd, 1H),7.08 (d, 2H), and 4.97 (s, 2H) ppm.

4.2.ad5-(N-Methyl-N-phenylsulfonylamino)-1,3-dihydro-1-hydroxy-2,1-benzoxaborole(C30)

M.p. 85-95° C. MS: m/z=304 (M+1) (ESI+) and m/z=302 (M−1) (ESI−). HPLC:96.6% purity at 254 nm and 89.8% at 220 nm. ¹H NMR (300 MHz, DMSO-d₆): δ9.23 (s, 1H), 7.72-7.63 (m, 2H), 7.56 (t, 2H), 7.50 (d, 2H), 7.16 (s,1H), 7.03 (d, 1H), 4.91 (s, 2H) and 3.14 (s, 3H) ppm.

4.2.ae 6-(4-Methoxyphenoxy)-1,3-dihydro-1-hydroxy-2,1-benzoxaborole(C31)

M.p. 126-129° C. MS: m/z=257 (M+1) (ESI+) and m/z=255 (M−1) (ESI−).HPLC: 98.4% purity at 254 nm and 98.4% at 220 nm. ¹H NMR (300 MHz,DMSO-d₆): δ 9.14 (s, 1H), 7.36 (d, 1H), 7.19 (s, 1H), 7.12 (d, 1H), 6.98(d, 2H), 6.95 (d, 2H), 4.93 (s, 2H) and 3.73 (s, 3H) ppm.

4.2.af 6-(4-Methoxyphenylthio)-1,3-dihydro-1-hydroxy-2,1-benzoxaborole(C32)

M.p. 95-100° C. MS: m/z=272 (M+), 273 (M+1) (ESI+) and m/z=271 (M−1)(ESI−). HPLC: 100% purity at 254 nm and 99.2% at 220 nm. ¹H NMR (300MHz, DMSO-d₆): δ 9.20 (s, 1H), 7.51 (d, 1H), 7.39-7.28 (m, 4H), 6.98 (d,2H), 4.93 (s, 2H) and 3.76 (s, 3H) ppm.

4.2.ag6-(4-Methoxyphenylsulfonyl)-1,3-dihydro-1-hydroxy-2,1-benzoxaborole(C33)

M.p. 180-192° C. MS: m/z=305 (M+1) (ESI+) and m/z=303 (M−1) (ESI−).HPLC: 96.8% purity at 254 nm and 95.5% at 220 nm. ¹H NMR (300 MHz,DMSO-d₆): δ 9.46 (s, 1H), 8.28 (s, 1H), 7.99 (d, 1H), 7.85 (d, 2H), 7.61(d, 1H), 7.11 (d, 2H), 5.02 (s, 2H) and 3.80 (s, 3H) ppm.

4.2.ah6-(4-Methoxyphenylsulfinyl)-1,3-dihydro-1-hydroxy-2,1-benzoxaborole(C34)

¹H NMR (300 MHz, DMSO-d₆): δ 9.37 (s, 1H), 8.02 (d, 1H), 7.71 (dd, 1H),7.59 (d, 2H), 7.53 (d, 1H), 7.07 (d, 2H), 5.00 (s, 2H) and 3.76 (s, 3H)ppm.

4.2.ai 5-Trifluoromethyl-1,3-dihydro-1-hydroxy-2,1-benzoxaborole (C35)

M.p. 113-118° C. MS: m/z=203 (M+1) (ESI+) and m/z=201 (M−1) (ESI−).HPLC: 100% purity at 254 nm and 100% at 220 nm. ¹H NMR (300 MHz,DMSO-d₆): δ 9.48 (s, 1H), 7.92 (d, 1H), 7.78 (s, 1H), 7.67 (d, 1H) and5.06 (s, 2H) ppm.

4.2.aj 4-(4-Cyanophenoxy)-1,3-dihydro-1-hydroxy-2,1-benzoxaborole (C36)

For coupling reaction between 4-fluorobenzonitrile and substitutedphenol to give starting material 2, see Igarashi, S.; et al. Chemical &Pharmaceutical Bulletin (2000), 48(11), 1689-1697.

¹H-NMR (300 MHz, DMSO-d₆) (ppm) 4.84 (s, 2H), 7.08 (d, J=8.2 Hz, 2H),7.18 (d, J=7.9 Hz, 1H), 7.45 (t, J=7.3 Hz, 1H), 7.63 (d, J=7.3 Hz, 1H),7.82 (d, J=8.5 Hz, 2H).

4.2.ak 5-(3-Cyanophenoxy)-1,3-dihydro-1-hydroxy-2,1-benzoxaborole (C37)

For coupling between 3-fluorobenzonitrile and substituted phenol to givestarting material 2: Li, F. et al., Organic Letters (2003), 5(12),2169-2171.

¹H-NMR (300 MHz, DMSO-d₆) (ppm) 4.93 (s, 2H), 7.0-7.1 (m, 2H), 7.3-7.4(m, 1H), 7.5-7.7 (m, 3H), 7.75 (d, J=8.2 Hz, 1H).

4.2.al 5-(4-Carboxyphenoxy)-1-hydroxy-2,1-benzoxaborole (C38)

To a solution of 5-(4-cyanophenoxy)-1-hydroxy-2,1-benzoxaborole obtainedin C17 (430 mg, 1.71 mmol) in ethanol (10 mL) was added 6 mol/L sodiumhydroxide (2 mL), and the mixture was refluxed for 3 hours. Hydrochloricacid (6 mol/L, 3 mL) was added, and the mixture was extracted with ethylacetate. The organic layer was washed with brine and dried on anhydroussodium sulfate. The solvent was removed under reduced pressure, and theresidue was purified by silica gel column chromatography (ethyl acetate)followed by trituration with diisopropyl ether to give the targetcompound (37 mg, 8%).

¹H-NMR (300 MHz, DMSO-d₆) δ (ppm) 4.94 (s, 2H), 7.0-7.1 (m, 4H), 7.76(d, J=7.9 Hz, 1H), 7.94 (d, J=8.8 Hz, 2H), 9.19 (s, 1H), 12.8 (br s,1H).

4.2.am 1-Hydroxy-5-[4-(tetrazole-1-yl)phenoxy]-2,1-benzoxaborole (C39)

A mixture of 5-(4-cyanophenoxy)-1-hydroxy-2,1-benzoxaborole (200 mg,0.797 mmol), sodium azide (103 mg, 1.59 mmol), and ammonium chloride (85mg, 1.6 mmol) in N,N-dimethylformamide (5 mL) was stirred at 80° C. fortwo days. Water was added, and the mixture was extracted with ethylacetate. The organic layer was washed with water and brine, and dried onanhydrous sodium sulfate. The solvent was removed under reducedpressure, and the residue was purified by silica gel columnchromatography (ethyl acetate) followed by trituration with ethylacetate to give the target compound (55 mg, 23%).

¹H-NMR (300 MHz, DMSO-d₆) δ (ppm) 4.95 (s, 2H), 7.0-7.1 (m, 2H), 7.23(d, J=8.8 Hz, 2H), 7.76 (d, J=7.9 Hz, 1H), 8.05 (d, J=8.5 Hz, 2H), 9.18(br s, 1H).

Example 5 Preparation of I from 2 Via 6

5.1 Catalytic Boronylation, Reduction and Cyclization

A mixture of 2 (10.0 mmol), bis(pinacolato)diboron (2.79 g, 11.0 mmol),PdCl₂(dppf) (250 mg, 3 mol %), and potassium acetate (2.94 g, 30.0 mmol)in 1,4-dioxane (40 mL) was stirred at 80° C. for overnight. Water wasadded, and the mixture was extracted with ethyl acetate. The organiclayer was washed with brine and dried on anhydrous sodium sulfate. Thesolvent was removed under reduced pressure. The crude product wasdissolved in tetrahydrofuran (80 mL), then sodium periodate (5.56 g,26.0 mmol) was added. After stirring at room temperature for 30 min, 2NHCl (10 mL) was added, and the mixture was stirred at room temperaturefor overnight. Water was added, and the mixture was extracted with ethylacetate. The organic layer was washed with brine and dried on anhydroussodium sulfate. The solvent was removed under reduced pressure, and theresidue was treated with ether to afford 6.3 mmol of the correspondingboronic acid. To the solution of the obtained boronic acid (0.595 mmol)in methanol (5 mL) was added sodium borohydride (11 mg, 0.30 mmol), andthe mixture was stirred at room temperature for 1 h. Water was added,and the mixture was extracted with ethyl acetate. The organic layer waswashed with brine and dried on anhydrous sodium sulfate. The solvent wasremoved under reduced pressure, and the residue was purified by silicagel column chromatography to give 0.217 mmol of I.

5.2 Results

Analytical data for exemplary compounds of structure I are providedbelow.

5.2.a 1,3-Dihydro-5-fluoro-1-hydroxy-2,1-benzoxaborole (C10)

Analytical data for this compound is listed in 4.2.j.

Example 6 Preparation of I from 3

6.1 One-Pot Boronylation and Cyclization

To a solution of 3 (4.88 mmol) and triisopropyl borate (1.35 mL, 5.86mmol) in tetrahydrofuran (10 mL) was added n-butyllithium (1.6 mol/L inhexanes; 6.7 mL, 10.7 mmol) dropwise over 15 min at −78° C. undernitrogen atmosphere, and the mixture was stirred for 2 h while allowingto warm to room temperature. The reaction was quenched with 2N HCl, andextracted with ethyl acetate. The organic layer was washed with brineand dried on anhydrous sodium sulfate. The solvent was removed underreduced pressure, and the residue was purified by silica gel columnchromatography and treated with pentane to give 0.41 mmol of I.

6.2 Results

Analytical data for exemplary compounds of structure I are providedbelow.

6.2.a 1,3-Dihydro-5-fluoro-1-hydroxy-2,1-benzoxaborole (C10)

Analytical data for this compound is listed in 4.2.j.

Example 7 Preparation of I from 3

7.1 One-Pot Boronylation and Cyclization with Distillation

To a solution of 3 (4.88 mmol) in toluene (20 mL) was added triisopropylborate (2.2 mL, 9.8 mmol), and the mixture was heated at reflux for 1 h.The solvent, the generated isopropyl alcohol and excess triisopropylborate were removed under reduced pressure. The residue was dissolved intetrahydrofuran (10 mL) and cooled to −78° C. n-Butyllithium (3.2 mL,5.1 mmol) was added dropwise over 10 min, and the mixture was stirredfor 1 h while allowing to warm to room temperature. The reaction wasquenched with 2N HCl, and extracted with ethyl acetate. The organiclayer was washed with brine and dried on anhydrous sodium sulfate. Thesolvent was removed under reduced pressure, and the residue was purifiedby silica gel column chromatography to give 1.54 mmol of I.

7.2 Results

Analytical data for exemplary compounds of structure I are providedbelow.

7.2.a 1,3-Dihydro-5-fluoro-1-hydroxy-2,1-benzoxaborole (C10)

Analytical data for this compound is listed in 4.2.j.

Example 8 Preparation of 8 from 7

8.1 Bromination

To a solution of 7 (49.5 mmol) in carbon tetrachloride (200 mL) wereadded N-bromosuccinimide (8.81 g, 49.5 mmol) and N,N-azoisobutylonitrile(414 mg, 5 mol %), and the mixture was heated at reflux for 3 h. Waterwas added, and the mixture was extracted with chloroform. The organiclayer was washed with brine and dried on anhydrous sodium sulfate. Thesolvent was removed under reduced pressure to give the crudemethyl-brominated intermediate 8.

Example 9 Preparation of 3 from 8

9.1 Hydroxylation

To crude 8 (49.5 mmol) were added dimethylformamide (150 mL) and sodiumacetate (20.5 g, 250 mmol), and the mixture was stirred at 80° C. forovernight. Water was added, and the mixture was extracted with ether.The organic layer was washed with water and brine, and dried onanhydrous sodium sulfate. The solvent was removed under reducedpressure. To the residue was added methanol (150 mL) and 1N sodiumhydroxide (50 mL), and the mixture was stirred at room temperature for 1h. The reaction mixture was concentrated to about a third of volumeunder reduced pressure. Water and hydrochloric acid were added, and themixture was extracted with ethyl acetate. The organic layer was washedwith water and brine, and dried on anhydrous sodium sulfate. The solventwas removed under reduced pressure, and the residue was purified bysilica gel column chromatography followed by trituration withdichloromethane to give 21.8 mmol of 3.

9.2 Results

Exemplary compounds of structure 3 prepared by the method above areprovided below.

9.2.a 2-Bromo-5-cyanobenzyl Alcohol

¹H-NMR (300 MHz, DMSO-d₆) δ ppm 4.51 (d, J=5.9 Hz, 2H), 5.67 (t, J=5.6Hz, 1H), 7.67 (dd, J=8.2, 2.0 Hz, 1H), 7.80 (s, J=8.2 Hz, 1H), 7.83 (d,J=2.0 Hz, 1H).

Additional examples of compounds which can be produced by this methodinclude 2-bromo-5-(4-cyanophenoxy)benzyl alcohol.

Example 10 Preparation of 9 from 2

10.1 Reaction

A mixture of 2 (20.0 mmol), (methoxymethyl)triphenylphosphonium chloride(8.49 g, 24.0 mmol), and potassium tert-butoxide (2.83 g, 24.0 mol) inN,N-dimethylformamide (50 mL) was stirred at room temperature forovernight. The reaction was quenched with 6 N HCl, and the mixture wasextracted with ethyl acetate. The organic layer was washed with water(×2) and brine, and dried on anhydrous sodium sulfate. The solvent wasremoved under reduced. To the residue were added tetrahydrofuran (60 mL)and 6 N HCl, and the mixture was heated at reflux for 8 h. Water wasadded, and the mixture was extracted with ether. The organic layer waswashed with brine and dried on anhydrous sodium sulfate. The solvent wasremoved under reduced pressure to afford 16.6 mmol of 9.

Example 11 Preparation Method of Step 13

11.1 Reaction

A solution of I in an appropriate alcohol solvent (R¹—OH) was refluxedunder nitrogen atmosphere and then distilled to remove the alcohol togive the corresponding ester.

Example 12 Preparation of Ib from Ia

12.1 Reaction

To a solution of Ia in toluene was added amino alcohol and theparticipated solid was collected to give Ib.

12.2 Results

(500 mg, 3.3 mmol) was dissolved in toluene (37 mL) at 80° C. andethanolamine (0.20 mL, 3.3 mmol) was added. The mixture was cooled toroom temperature, then ice bath, and filtered to give C40 as a whitepowder (600.5 mg, 94%).

12.2a (C40)

¹H-NMR (300 MHz, DMSO-d₆) δ (ppm) 2.88 (t, J=6.2 Hz, 2H), 3.75 (t, J=6.3Hz, 2H), 4.66 (s, 2H), 5.77 (br, 2H), 6.85-6.91 (m, 2H), 7.31 (td,J=7.2, 1.2 Hz, 1H).

Example 13

Formulations

Compounds of the present invention can be administered to a patientusing a therapeutically effective amount of a compound of Formulae (I)or (II) in any one of the following three lacquer formulations and onesolvent formulation. The lacquer formulation provides good durabilitywhile the solvent formulation provides good ease of use. These compoundscan also be applied using a spray formulation, paint-on lacquer, drops,or other.

-   -   1. 20% propylene glycol; 70% ethanol; 10% compound of invention;    -   2. 70% ethanol; 20% poly(vinyl methyl ether-alt-maleic acid        monobutyl ester); 10% compound of the invention;    -   3. 56% ethanol; 14% water; 15% poly(2-hydroxyethyl        methacrylate); 5% dibutyl sebacate; 10% compound of the        invention;    -   4. 55% ethanol; 15% ethyl acetate; 15% poly(vinyl acetate); 5%        dibutyl sebacate; 10% compound of the invention.

The preparation of these formulations is well known in the art and isfound in references such as Remington: The Science and Practice ofPharmacy, supra.

Example 14

Antifungal MIC Testing

All MIC testing followed the National Committee for Clinical LaboratoryStandards (NCCLS) guidelines for antimicrobial testing of yeasts andfilamentous fungi (Pfaller et al., NCCLS publication M38-A—ReferenceMethod for Broth Dilution Antifungal Susceptibility Testing ofFilamentous Fungi; Approved Standard. Wayne, Pa.: NCCLS; 2002 (Vol. 22,No. 16) except the Malassezia species which was incubated in a ureabroth (Nakamura et al., Antimicrobial Agents And Chemotherapy, 2000,44(8) p. 2185-2186). Results of the MIC testing is provided in FIGS.1A-1C.

Example 15

Keratin Assay

Many antifungal agents strongly bind to keratin which not only reducestheir antifungal potency but also may restrict their penetration intothe nail. The affinities of the compounds for keratin powder wasdetermined by a method described in Tatsumi, Antimicrobial Agents andChemotherapy, 46(12):3797-3801 (2002).

A comparison of MIC data for several compounds of the invention againstT. rubrum, with and without the presence of 5% keratin, is provided inFIGS. 1A-1C.

Example 16

(C10) Antifungal Spectrum of Activity

(C10) is a novel compound in development for use as a topical antifungaltreatment. The purpose of this study was to determine the minimuminhibitory concentration (MIC) for (C10) against 19 test strains offungi including: Aspergilus fumigatus (A. fumigatus), Candida Albicans(C. albicans, both fluconazole sensitive and resistant strains), Candidaglabrata (C. glabrata), Candida krusei (C. krusei), Cryptococcusneoformans (C. neoformans), Candida parapsilosis (C. parapsilosis),Candida tropicalis (C. tropicalis), Epidermophyton floccosum (E.floccosum), Fusarium solani (F. solani), Malassezia furfur (M. furfur),Malassezia pachydermatis (M. pachydermatis), Malassezia sympodialis (M.sympodialis), Microsporum audouinii (M. audouinii), Microsporum canis(M. canis), Microsporum gypseum (M. gypseum), Trichophytonmentagrophytes (T. mentagrophytes), Trichophyton rubrum (T. rubrum),Trichophyton tonsurans (T. tonsurans). Fungal growth was evaluated afterexposure to different concentrations of (C10). In addition, the MIC for(C10) against T. rubrum in the presence of 5% keratin powder and theminimum fungicidal concentration (MFC) for (C10) against T. rubrum andT. mentagrophytes were also determined. Ciclopirox and/or terbinafineand/or fluconazole and/or itraconazole were used as comparators andtested in a similar manner. These studies were conducted at NAEJAPharmaceutical, Inc.

Materials and Methods

(C10) was obtained from Anacor Pharmaceuticals, Inc. (Palo Alto, Calif.,USA). ATCC strains were obtained from ATCC (Manassas, Va., USA).Ciclopirox-olamine was obtained from Sigma-Aldrich Co. (St. Louis, Mo.,USA). Terbinafine, fluconazole and itraconazole were synthesized atNAEJA Pharmaceutical Inc. (Edmonton, AB, Canada), experimentalprocedures and analytical data for these standards are stored in NAEJAarchives.

All MIC testing followed the National Committee for Clinical LaboratoryStandards (NCCLS) guidelines for antimicrobial testing of yeasts andfilamentous fungi (Pfaller et al., 2002) except the Malassezia specieswhich were incubated in a urea broth (Nakamura et al., 2000). Themicrobroth dilution method was used to test the in vitro activity of(C10) against 19 test strains of fungi. Briefly, compounds weredissolved in DMSO and diluted in sterile water to give a working stock.Two-fold serial dilutions of the working stock were prepared in 96-wellplates and media was added. Media was RPMI, RPMI+MOPS, modified RPMI, ormodified Urea broth. The plates were inoculated with the fungalsuspensions to give a final inoculum size of 0.5-2.5×10³ cells/mL foryeasts or 0.4-5×10⁴ CFU/mL for filamentous fungi and then incubated for24-168 h at 35° C. The final concentration of DMSO did not exceed 5%.The MIC was defined as the lowest concentration that resulted in over90% reduction of growth, as compared to a drug-free control. The MFC wasdefined as the lowest concentration that killed over 90% of the fungi,as compared to a drug-free control.

Results and Conclusions

The results for the MIC of (C10) and reference compounds against 19strains of fungi are shown in FIG. 2. The results for the MFC of AN2690against 2 strains of fungi are shown in Table 2. (C10) had MIC valuesranging from 0.25-2 μg/mL against all fungi tested. Addition of 5%keratin powder to the media did not effect the MIC against T. rubrum.(C10) had fungicidal activity against T. rubrum and T. mentagrophyteswith MFC values of 8 and 16 μg/mL, respectively. Reference compounds hadMIC values in the range defined by NCCLS.

Example 17

The Solubility, Stability and Log P Determination of Compounds of thePresent Invention by LC/MS/MS

The solubility, room temperature stability and Log P of C10 wasdetermined by the following methodology.

Reagents and Standards:

Ethanol: 200 proof ACS Grade (EM Science, Gibbstown, N.J., USA);Octanol: Octyl alcohol (EM Science, Gibbstown, N.J., USA); Acetonitrile:HPLC Grade (Burdick & Jackson, Muskegon, Mich., USA); Ammonium Acetate:lot 3272×49621 (Mallinckrodt, Phillipsburg, N.J., USA); C10: lotA032-103 (Anacor Pharmaceuticals, Palo Alto, Calif., USA); p-Nitrophenol(PNP): lot OGNO1 (TCI America, Portland, Oreg., USA); Water: Deionizedwater (from Millipore systems, Billerica, Mass., USA)

Solubility

N-Octanol and water were mutually pre-saturated by vigorously stirring amixture of both solvents for up to 12 h and the mixture was allowed toseparate. Solubility in each solvent was determined by adding 10 μL of20, 40, 200, 1000 and 5000 μg/mL of C10 in DMSO to the pre-saturatedn-octanol or water. After the sample was vortexed for 10 sec, the samplewas centrifuged for 10 min at ca. 3000 rpm. A visual inspection was madeto determine if the sample was clear or if a pellet had formed on thebottom of the tube.

Log P

C10 (10 μL of 5000 μ/mL) at 2× the final concentration was added to 0.5mL pre-saturated n-octanol and mixed. An equal volume (0.5 mL) ofpre-saturated water was added, vortex mixed and then mixed on a rotatingshaker for one hour and 24 h in triplicate at ca. 25° C. The organic andaqueous layers were separated by centrifugation for 5 min at ca. 2000rpm. Twenty five μL of the octanol (top) layer were removed and placedin a pre-labeled tube. Twenty five μL of the aqueous layer (bottom) wereremoved, taking care to avoid octanol contamination, and placed in apro-labeled tube.

Stability at Room Temperature

C10 (10 μL of 5000 μg/mL) was added both to 0.5 mL n-octanol and 0.5 mLwater in triplicate. Samples were mixed. At 0 h and 24 h samples werestored at ca. −20° C. Twenty five μL of sample was used for analysis.

Extraction Procedure C10

For the octanol sample, 25 μL of ethanol, 25 μL of water and 300 μL ofacetonitrile containing the internal standard was added. For the watersample, 25 μL of ethanol, 25 μL of octanol and 300 μL of acetonitrilecontaining the internal standard [60 mL of acetonitrile add 6 μL of PNP(1000 μg/mL)] was added. For the calibrators 25 μL of octanol, 25 μL ofwater and 300 pL of acetonitrile containing the internal standard wasadded. The sample was vortexed for 10 seconds. Two hundred μL of theorganic layer were transferred into a clean deactivated autosamplervial.

Calculations

A 1/concentration weighted linear regression was used for thequantitation of C10. All integration were performed with peak areasusing Analyst version 1.3, Applied Biosystems. For C10, peak area ratiosanalyte to internal standard PNP were used for all quantitation.

The partition coefficient (P) was calculated according to the equationdetailed below:P=[Sample concentration]_(octanol)/[Sample concentration]_(water)Log P=log₁₀(partition coefficient)Results:

As shown in Table 17A the solubility of C10 in both octanol and water isvery good over the concentration range tested.

TABLE 17A Solubility of C10 in water and octanol Targeted Conc WaterOctanol (μg/mL) Visual Visual 0.800 Clear Clear 4.00 Clear Clear 20.0Clear Clear 100 Clear Clear

Table 17B shows the results of the log P determination after 1 h and 24h for C10. The mean log P after 1 h was 1.97 (n=3). After 24 h theconcentrations in both the octanol and water layer remained the same.The mean log P after 24 h was 1.93 (n=3).

TABLE 17B Log P of C10 Conc. in Water Conc. in Octanol Sample (μg/mL)(μg/mL) Log P 1 h-1 1.26 108 1.93 1 h-2 1.21 103 1.93 1 h-3 1.05 1152.04 24 h-1 1.27 104 1.91 24 h-2 1.17 109 1.97 24 h-3 1.28 99.0 1.89

A stability study for C10 was initiated at room temperature over 24 hwithout continuous mixing. Table 17C shows that C10 in pure water andoctanol is stable over 24 h.

TABLE 17C Water and Octanol stability for C10 at room temperature after24 h. Percent Mean Remaining 24 h Sample (μg/mL) SD versus 0 g Water-0 h82.5 3.72 115 Water-24 h 95.0 21.4 Octanol-0 h 115 3.06 93 Octanol-24 h107 6.11

Example 18

Determination of Penetration of C10 into the Human Nail

Two nail penetration studies were performed based on the protocol in Huiet al., Journal of Pharmaceutical Sciences, 91(1): 189-195 (2002) (“Huiprotocol”). The purpose of this study was to determine and compare thepenetration and distribution of C10 in vehicle into the human nail platein vitro relative to 8% ciclopirox w/w in commercial lacquer (Penlac®).

Materials and Methods

Test Article and Dosage Formulation

8% ciclopirox w/w in commercial lacquer was manufactured by Dermick(Berwyn, Pa.). The radiochemical purity and specific activity of thechemical was determined as >95% and 12.5 mCi/mmol, respectively.

The study was composed of two groups. The compositions (weight %) of thedosage formulations are as follows:

Active radiolabeled compound in four groups.

Dosing Test Chemical Radioactivity Groups* (×14 days) (%) (per 10 μL) A(C10) qd 10 0.19 μCi C (Ciclopirox) qd 8 0.22 μCi *A = C10 group, C =Ciclopiriox group

Human Nails

Healthy human finger nail plates were collected from adult humancadavers and stored in a closed container at 0-4° C. Before theexperiment, the nail plates were gently washed with normal saline toremove any contamination, then re-hydrated by placing them for threehours on a cloth wetted with normal saline. The nail samples wererandomly selected into four groups.

Dosing and Surface Washing Procedures

Dose Preparation:

Radioactivity of each group is approximately 0.19±0.01 and 0.22±0.03μCi/10 μL solutions respectively, for ¹⁴C—C10 (group A), and¹⁴C-ciclopirox (group C).

Experiment Procedure:

Study Group A Group C Day wash dose sample wash dose sample 1 D D 2 W DW D 3 W D C W D C 4 W D W D 5 W D W D 6 W D C W D C 7 W D W D 8 W D W D9 W D C W D C 10 W D W D 11 W D W D 12 W D C W D C 13 W D W D 14 W D W D15 W C, N W C, N W = once per day before dosing (9~10 AM). D = once perday (9~10 AM). C = changing/sampling cotton ball after surface washingbefore topical dosing. N = Nail sampling.Washing Procedure

Surface washing was started in morning 10 min prior to next dosing, thesurface of the nail was washed with cotton tips in a cycle, as follows:

a tip wetted with absolute ethanol, then

a tip wetted with absolute ethanol, then

a tip wetted with 50% IVORY liquid soap, then

a tip wetted with distilled water, then

a final tip wetted with distilled water.

The washing samples from each cycle of each nail were pooled andcollected by breaking off the cotton tip into scintillation glass vials.Aliquots of 3.0 mL methanol were added into each vial to extract testmaterial. The radioactivity of each sample was measured in a liquidscintillation counter.

Incubation System

A Teflon one-chamber diffusion cell (PermeGear, Inc., Hellertown, Pa.)was used to hold each nail. To approximate physiological conditions, asmall cotton ball wetted with 0.1 mL normal saline was placed in thechamber to serve as a nail bed and provide moisture for the nail plate.Every 3 days, 0.1 mL normal saline was injected through the inlet intothe chamber to keep the cotton ball wet. The nail plate was placed on aledge inside the receptor (1.0 cm in diameter and 0.5 cm high). Theventral (inner) surface of the nail was placed face down and rested onthe wet cotton ball. The cells were placed on a platform in a largeglass holding tank filled with saturated sodium phosphate solution tokeep the cells at a constant humidity of 40%.

Sampling Instrument

The nail sampling instrument had two parts, a nail sample stage and adrill. The nail sampling stage consists of a copper nail holder, threeadjustments, and a nail powder capture. Three adjustments allow movementin vertical direction. The first coarse adjustment (on the top) was forchanging the copper cell and taking powder samples from the capture. Theother two adjustments (lower) were for sampling process. The secondcoarse adjustment allowed movement of 25 mm and the fine adjustmentprovides movement of 0.20 mm. The nail powder capture was locatedbetween the copper cell and the cutter. The inner shape of the capturewas inverted funnel and the end of funnel connects to a vacuum. Byplacing a circle filter paper inside of the funnel, the nail powdersamples were captured on the filter paper during the sampling process.

Sampling Procedure

After completion of the incubation phase, the nail plate was transferredfrom the diffusion cell to a clean copper nail holder for samplingprocess. The nail plate was inverted so that the ventral (nail bed)surface now faced up and the dorsal (outer) dosed surfaced faced down.The copper nail holder has an opening as it sits on top of the stage.When the sampling process initiated, the coarse adjustment was adjustedto move the position of the stage until the nail plate was just touchingthe tip of the cutter. Then the drill was turned on and the fineadjustment was turned to push the stage closer to the drill, removing anail core sample. After the above process, approximate 0.40-0.50 mm indepth and 7.9 mm in diameter nail pulverized samples were harvested fromthe center of the ventral (nail bed) surface of the nail.

The powdered nail samples were collected into a glass scintillation vialand weighted. Aliquots of 5.0 mL Packard soluene-350 (Packard InstrumentCompany, Meriden, Conn.) was added to the scintillation vial to dissolvethe powder. The upper part, the intermediate and dorsal layers of thecenter of the nail, including the area of application of the dose wascut in the same diameter as the sampled area and was then placed into aglass scintillation vial with 5.0 mL packard soluene-350. The rest ofthe nail was also placed in a glass scintillation vial with 5.0 mLpackard soluene-350.

The amount of nail sample removed was measured by the difference inweight of the nail plate before and after drilling, and collecting thecore of powder.

Radioactivity Measurement

All radioactivity measurements were conducted with a Model 1500 LiquidScintillation Counter (Packard Instrument Company, Downer Grove, Ill.).The counter was audited for accuracy using sealed samples of quenchedand unquenched standards as detailed by the instrument manual. The ¹⁴Ccounting efficiency is equal to or greater than 95%. All nail samplespre-treated with packard soluene-350 were incubated at 40° C. for 48hours followed by the addition of 10 mL scintillation cocktail(HIONIC-FLUOR, Packard Instrument Company, Meriden, Conn.). Othersamples (standard dose, surface washing, and bedding material) weremixed directly with Universal ES scintillation cocktail (ICNBiomedicals, Costa Mesa, Calif.). Background control and test sampleswere counted for 3 minutes each for radioactivity.

Data Analysis

All sample counts (expressed as dpm) were transcribed by hand to acomputerized spreadsheet (Microsoft Excel). The individual and mean(±S.D.) amount of test chemical equivalent in nail, bedding material,and wash samples are presented as dpm, μCi, percent administered dose,and mg equivalent at each time point. The concentration of ¹⁴C-labeledtest chemicals were calculated from the value based on the specificactivity of each [¹⁴C]-test chemical. The information of concentrationof non-labeled test chemical in the topical formulation was obtainedfrom the manufactures. Total concentration of test chemical equivalentis the sum of the concentration of ¹⁴C-labeled test chemical and theconcentration of non-labeled test chemical. The value of total amount oftest chemical equivalent in each nail sample was calculated from thosevalues based on radioactivity of the sample and the ratio of total mgtest chemical equivalent and radioactivity of the test chemical. Thedata was further normalized by dividing with the weight of the sample.Statistical significant of nail samples from every two groups wasanalyzed by student t-test.

Terminology

Ventral/intermediate center: Powdered nail sample drilled from thecenter of the inner surface (facing the nail bed) approximately 0.3-0.5mm in depth to the surface. The area is beneath the dosed site of thenail place but does not include dosed surface (dorsal nail surface).

Dorsal/intermediate center: Immediate area of dosed site.

Remainder nail: The remaining part of the nail that has not been dosed.

Supporting bed: The cotton ball placed within the Teflon chamber of thediffusion cell to provide moisture to the nail plate and also to receivechemicals penetrating through the nail plate.

Surfacing washing: Ethanol (or other organic solvents) and soap/waterwashing on the surface of the dosed site.

Ring: A plastic ring placed on the top of the nail plate to preventleakage from the dose site onto rest of the nail plate or inside of thecell chamber.

Cell washing: Ethanol (or other organic solvents) and soap/water wash ofthe inside of the diffusion cell.

Results

Characteristics of Nail Samples

For both groups (Group A group and Group C) the thickness of whole nailplate, the depth of the ventral surface core sample removed by cutter,the percentage of the whole nail thickness, and the actual weight ofpowdered nail sample were collected. No statistical difference is foundbetween two groups (P>0.05).

Weight Normalized C10 and Ciclopirox Equivalent in Nail

FIG. 3 shows summarized normalized drug equivalents in each part (layer)of nail samples. After weight normalization, the concentration of C10equivalent in dorsal/intermediate center, ventral/intermediate center,and remainder nail samples was significantly higher than that ofciclopirox equivalent (p≦0.002).

C10 and Ciclopirox Equivalent in Cotton Ball Nail Supporting Bed

FIG. 4 shows summarized C10 and ciclopirox equivalent in supporting bedcotton ball samples. Similar to weight normalized C10 equivalent in thenail plate samples, absolute amount of C10 equivalent per cotton ballsample in group A (after 14 day dosing) was significantly higher thanthat of ciclopirox in group C (p≦0.004). The difference of these twotest chemicals was 250 times.

Mass Balance of Radioactivity of [¹⁴C]-C10 and [¹⁴C]-Ciclopirox after14-day Treatment

Table 5 shows summarized radioactive recovery from washing, nailsamples, and supporting bed cotton ball samples. Cumulativeradioactivity recoveries of carbon-14 were 88±9.21, and 89±1.56 percentof applied dose in group A, and group C, respectively. 88% of theradiolabeled material was accounted for.

Conclusion

In this study, penetration rate of [¹⁴C]-C10 in Anacor topicalformulation and [¹⁴C]-ciclopirox (8% w/w in commercial lacquer) intohuman nail with four different dosing and washing methods was studied.

Results show that much more amount of [¹⁴C]-C10 penetrating into thedeeper parts of the nail when compared with [¹⁴C]-ciclopirox. Tables 3and 4 show that the amount of [¹⁴C]-C10 equivalent inventral/intermediate center of the nail layer and cotton ball supportingbed in the group A was statistically higher (p≦0.002) than group C aftera 14-day dosing period.

Example 19

Determination of Penetration of C10 into the Human Nail

The aim of the current study was to assess and compare the perungualabsorption of C10 in a simple vehicle using MedPharm's TurChub® model(see http://www.medpharm.co.uk; specificallyhttp://www.medpharm.co.uk/downloads/Skin%20and%20nail%20dec %202003.pdf;viewed Feb. 14, 2006). in a full scale experiment. Six replicatesinvolving C10 were conducted and Formulations Y (8% ciclopirox w/w incommercial lacquer) and Z (Loceryl, 5% amorolfine w/v in commerciallacquer) were used as the reference formulations.

The following materials were used in these experiments. These materialswere used without any modifications.

A dose of 40 μL/cm² of the test compound C10 in 50:50 propyleneglycol:ethyl acetate was applied to a full thickness nail sample eachday over a total duration of five days. Both the reference formulationswere also applied at the same dose.

TurChub® Zone of Inhibition Experiment

Placebo, test item C10 in vehicle and the reference formulations Y and Zwere tested for their inhibition of Trichophyton rubrum (T. rubrum)growth after penetration through a full thickness human nail using azone of inhibition measurement.

Formulation Efficacy Testing

FIGS. 5-9 show the results obtained from the TurChub zone of inhibitionassays. It can be observed that C10 is a potent antifungal agent, whichcan penetrate through a full thickness nail to elicit its effect againstthe target organism T. rubrum. No zones of inhibition were observed withreference formulations Y and Z or with the placebo for C10. Theexperiment using C10 was repeated for a second time to confirm theresult and it can be observed from FIGS. 6 and 7 that C10 shows zones ofinhibition of 100%, 67%, 46%, 57%, 38% and 71% in the first experimentand 74%, 86%, 100%, 82%, 100% and 84% in the second experiment. Themeasurement was taken from the nail to the first point of growthobserved.

From the results obtained using MedPharm's TurChub zone of inhibitionassay as a test system, the test item C10 was found to be a powerfulantifungal agent and demonstrated superior results vs. the commercialreference formulations Y and Z. From these experiments it appears thatthe compound is permeating through a full thickness nail barrier toexhibit the antifungal activity.

Example 20

Determination of Penetration of C10 into the Human Nail: Dose Response

The optimal dose-response range for penetration into the human nail wasdetermined to be between 1% and 15%. The experiments to determine theoptimal dose-response was conducted as follows.

Tests at different test compound concentrations were conducted on nailsderived from the same cadaver. Cadaver nails were hydrated overnight,cut into 4 equally sized squares and placed onto individual poloxomersupports. Test articles were formulated in a lacquer at 1%, 2.5%, 5%,7.5%, 10% and 15% w/v. A 40 μL/cm² dose is applied to the center of thenail piece and the nails are left for 24 hrs. Nails are removed from thepoloxomer support. Poloxomer support is analyzed for quantity ofcompound using LC/MS/MS.

It is understood that the examples and embodiments described herein arefor illustrative purposes only and that various modifications or changesin light thereof will be suggested to persons skilled in the art and areto be included within the spirit and purview of this application andscope of the appended claims. All publications, patents, and patentapplications cited herein are hereby incorporated by reference in theirentirety for all purposes.

What is claimed is:
 1. A pharmaceutical composition, comprising: (a)1,3-dihydro-5-chloro-1-hydroxy-2,1-benzoxaborole, or a pharmaceuticallyacceptable salt thereof; (b) a pharmaceutically acceptable excipient foruse in an animal suffering from an infection by a microorganism.
 2. Thepharmaceutical composition of claim 1, wherein said pharmaceuticalcomposition is for topical administration.
 3. The pharmaceuticalcomposition of claim 1, wherein said excipient is a pharmaceuticallyacceptable topical carrier.
 4. The pharmaceutical composition of claim1, wherein said formulation is a member selected from a lacquer, lotion,cream, gel, ointment, and spray.
 5. The pharmaceutical composition ofclaim 1, wherein said composition is a lacquer.
 6. The pharmaceuticalcomposition of claim 1, wherein said composition is a gel.
 7. Thepharmaceutical composition of claim 1, wherein said composition furthercomprises one or more members selected from an emulsifier, emollient,antioxidant, preservative, chelating agent, neutralizing agent,viscosity increasing agent, penetration enhancer, anti-inflammatoryagent, vitamin, anti-aging agent, sunscreen, and acne-treating agent. 8.The pharmaceutical composition of claim 1, wherein said compositionfurther comprises a member selected from the group consisting of athickener, a gel phase carrier, nail penetration enhancer, and aviscosity increasing agent.
 9. The pharmaceutical composition of claim1, wherein said composition comprises a chelating agent.
 10. Thepharmaceutical composition of claim 9, wherein said chelating agent isselected from the group selected from citric acid, ethylene diaminetetraacetic acid (EDTA), ethylene glycol-bis(beta-aminoethylether)-N,N,N′,N′-tetraacetic acid (EGTA) and8-Amino-2-[(2-amino-5-methylphenoxy)methyl]-6-methoxyquinoline-N,N,N′,N′-tetraaceticacid, tetrapotassium salt (QUIN-2).
 11. The pharmaceutical compositionof claim 9, wherein said chelating agent is ethylene diamine tetraaceticacid.
 12. The pharmaceutical composition of claim 9, wherein saidchelating agent is present in an amount of between 0.005% to 2% byweight.
 13. The pharmaceutical composition of claim 1, wherein saidcomposition comprises alcohol.
 14. The pharmaceutical composition ofclaim 1, wherein said composition comprises alcohol and water.
 15. Thepharmaceutical composition of claim 1, wherein said compositioncomprises one or more members selected from ethanol and propyleneglycol.
 16. The pharmaceutical composition of claim 1, wherein said1,3-dihydro-5-chloro-1-hydroxy-2,1-benzoxaborole is present in saidcomposition in a concentration from about 0.5% to about 15% w/v.
 17. Thepharmaceutical composition of claim 1, wherein said1,3-dihydro-5-chloro-1-hydroxy-2,1-benzoxaborole is present in saidcomposition in a concentration from about 0.1% to about 12.5% w/v. 18.The pharmaceutical composition of claim 1, wherein said1,3-dihydro-5-chloro-1-hydroxy-2,1-benzoxaborole is present in saidcomposition in a concentration from about 1% to about 5% w/v.
 19. Thepharmaceutical composition of claim 1, wherein said1,3-dihydro-5-chloro-1-hydroxy-2,1-benzoxaborole is present in saidcomposition in a concentration from about 2% to about 8% w/v.
 20. Thepharmaceutical composition of claim 1, wherein said1,3-dihydro-5-chloro- 1-hydroxy-2,1-benzoxaborole is present in saidcomposition in a concentration from about 4% to about 9% w/v.
 21. Thepharmaceutical composition of claim 1, wherein said1,3-dihydro-5-chloro-1-hydroxy-2,1-benzoxaborole is present in a formwhich is a member selected from a hydrate with water, a solvate with analcohol, an adduct with an amino compound, and an adduct with an acid.22. The pharmaceutical composition of claim 1, wherein a site of saidtopical administration is skin or nail or hair or skin surrounding thenail or skin surrounding the hair.
 23. The pharmaceutical composition ofclaim 1, wherein the microorganism is a fungus or a yeast.
 24. Thepharmaceutical composition of claim 23, wherein said fungus or yeast isa member selected from Candida species, Trichophyton species,Microsporium species, Aspergillus species, Cryptococcus species,Blastomyces species, Cocciodiodes species, Histoplasma species,Paracoccidiodes species, Phycomycetes species, Malassezia species,Fusarium species, Epidermophyton species, Scytalidium species,Scopulariopsis species, Alternaria species, Penicillium species,Phialophora species, Rhizopus species, Scedosporium species andZygomycetes species.
 25. The pharmaceutical composition of claim 23,wherein said fungus or yeast is a member selected from Aspergilusfumigatus, Blastomyces dermatitidis, Candida albicans, Candida glabrata,Candida krusei, Cryptococcus neoformans, Candida parapsilosis, Candidatropicalis, Cocciodiodes immitis, Epidermophyton floccosum, Fusariumsolani, Histoplasma capsulatum, Malassezia furfur, Malasseziapachydermatis, Malassezia sympodialis, Microsporum audouinii,Microsporum canis, Microsporum gypseurn, Paracoccidiodes brasiliensis,Trichophyton mentagrophytes, Trichophyton rubrum and Trichophytontonsurans.
 26. The pharmaceutical composition of claim 23, wherein saidfungus or yeast is a member selected from Trichophyton concentricum,Trichophyton violaceum, Trichophyton schoenleinii, Trichophytonverrucosum, Trichophyton soudanense, Microsporum gypseum, Microsporumequinum, Candida guilliermondii, Malassezia globosa, Malassezia obtuse,Malassezia restricta, Malassezia slooffiae and Aspergillus flavus. 27.The pharmaceutical composition of claim 23, wherein said fungus or yeastis a dermatophyte.
 28. The pharmaceutical composition of claim 23,wherein said fungus or yeast is a member selected from Tinea unguium,Trichophyton rubrum and Trichophyton mentagrophytes.
 29. Thepharmaceutical composition of claim 1, wherein the infection is acutaneous infection.
 30. The pharmaceutical composition of claim 1,wherein the infection is a member selected from an ungual, periungualand subungual infection.
 31. The pharmaceutical composition of claim 1,wherein the infection is onychomycosis.
 32. The pharmaceuticalcomposition of claim 1, wherein the animal is a human.
 33. Thepharmaceutical composition of claim 1, wherein said composition is in acosmetically effective amount.
 34. The pharmaceutical composition ofclaim 1, wherein said composition is in a therapeutically effectiveamount.
 35. A method of treating an infection in an animal, said methodcomprising administering to the animal a therapeutically effectiveamount of 1,3-dihydro-5-chloro-1-hydroxy-2,1-benzoxaborole, or apharmaceutically acceptable salt thereof, sufficient to treat saidinfection.
 36. The method of claim 35, wherein said infection is amember selected from a systemic infection, a cutaneous infection, and anungual or periungual infection.
 37. The method of claim 35, wherein saidinfection is a member selected from chloronychia, paronychia,erysipeloid, gonorrhea, swimming-pool granuloma, leprosy, acutebacterial perionyxis, sporotrichosis, syphilis, tuberculosis verrucosacutis, tularemia, Mycotic keratitis, Extension oculomycosis, Endogenousoculomycosis, Lobomycosis, Mycetoma, Piedra, Pityriasis versicolor,Tinea corporis, Tinea cruris, Tinea pedis, Tinea barbae, Tinea capitis,Tinea nigra, Otomycosis, Tinea favosa, Chromomycosis, and Tineaimbricata.
 38. The method of claim 35, wherein said infection isonychomycosis.
 39. The method of claim 35, wherein said onychomycosis istinea unguium.
 40. The method of claim 35, wherein said animal is amember selected from a cattle, goat, pig, sheep, horse, cow, bull, dog,guinea pig, gerbil, rabbit, cat, chicken, and turkey.
 41. The method ofclaim 35, wherein said animal is a human.
 42. The method of claim 35,wherein, the administering is at a site which is a member selected fromthe skin, nail, hair, hoof and claw.
 43. The method of claim 42, whereinsaid skin is the skin surrounding the nail, hair, hoof or claw.
 44. Themethod of claim 35, wherein said infection is a fungal infection.
 45. Amethod of treating onychomycosis in a human, said method comprisingadministering to the human a therapeutically effective amount of1,3-dihydro-5-chloro-1-hydroxy-2,1-benzoxaborole, or a pharmaceuticallyacceptable salt thereof, sufficient to treat said onychomycosis.
 46. Amethod of inhibiting the growth of a fungus in a human, said methodcomprising administering to the human a therapeutically effective amountof 1,3-dihydro-5-chloro-1-hydroxy-2,1-benzoxaborole, or apharmaceutically acceptable salt thereof.